Antisense RNA standardizing control

ABSTRACT

Methods for producing a population of distinct aRNA molecules from an initial population of distinct mRNA molecules are provided. In the subject methods, an initial mRNA sample is contacted with a population of distinct tagged antisense molecules to produce a population of hybrid or duplex molecules. The resultant population of hybrid molecules, or template derivatives thereof, is then contacted with a DNA dependent RNA polymerase to produce the population of distinct aRNA molecules. Also provided are kits for practicing these methods. The subject methods find use a variety of different applications in which the preparation of aRNA is desired, e.g., the preparation of nucleic acid targets for use in array based hybridization applications.

BACKGROUND OF THE INVENTION

[0001] The technical field of this invention is molecular biology, andparticularly tools for nucleic acid array standardization.

[0002] The characterization of cell specific gene expression findsapplication in a variety of disciplines, such as in the analysis ofdifferential expression between different tissue types, different stagesof cellular growth or between normal and diseased states. Fundamental tothe characterization of cell specific gene expression is the detection,qualitative or quantitative, of mRNA. However, the detection of mRNA isoften complicated by one or more of the following factors: cellheterogeneity, paucity of material, or limits of low abundance mRNAdetection.

[0003] One method which has been developed to address at least some ofthe problems associated with mRNA detection is known as “antisense RNA”(aRNA) amplification. In this method, first strand cDNA is prepared frommRNA using an oligo dT primer that comprises an RNA polymerase promoterlocated at the 5′ end of the oligo dT region. The first strand cDNA isthen converted to ds cDNA. Finally, the ds cDNA is contacted with theappropriate RNA polymerase under conditions sufficient to produce aRNA.The method can be adjusted to obtain amplification of the initial mRNAof up to 106 fold. The aRNA can then be used in a variety ofapplications as hybridization target, for cDNA library construction andthe like, where such applications include assays for differential geneexpression.

[0004] Current methods of antisense RNA amplification as described abovethat employ RNA intermediates are not entirely satisfactory. Forexample, methods currently employed require the synthesis of one or moreDNA strands, e.g., first and second cDNA, in addition to RNAtranscription with the RNA polymerase, and therefore require multiplesteps using multiple reagents.

[0005] Accordingly, there is interest in the development of improvedmethods of antisense RNA amplification which do not suffer from one ormore of the above deficiencies experienced using current methods.

[0006] U.S. patents disclosing methods of antisense RNA synthesisinclude: U.S. Pat. Nos. 6,312,928; 6,309,384; 6,132,997; 5,932,451;5,869,249; 5,716,785; 5,593,863; 5,554,516; 5,545,522; 5,514,545;5,512,462; 5,470,724; 5,437,990; 5,399,491; 5,130,238; 5,021,335; and4,683,195. Antisense RNA synthesis is also discussed in Phillips &Eberwine, Methods: A Companion to Methods in Enzymology (1996)10:283-288; Eberwine et al., Proc. Natl. Acad. Sci. USA (1992) 89:3010-3014; Eberwine, Biotechniques (1996) 20:584-591; and Methods inEnzymology (1992) 216:80-100.

SUMMARY OF THE INVENTION

[0007] Methods for producing a population of distinct aRNA moleculesfrom an initial population of distinct mRNA molecules are provided. Inthe subject methods, an initial mRNA sample is contacted with apopulation of distinct tagged antisense nucleic acid molecules toproduce a population of hybrid molecules. The resultant hybridmolecules, or derivatives thereof (e.g., template structures producedthere from) are then transcribed into aRNA molecules using a DNAdependent RNA polymerase transcription step Also provided are kits forpracticing these methods. The subject methods find use a variety ofdifferent applications in which the preparation of aRNA is desired,e.g., the preparation of nucleic acid targets for use in array basedhybridization applications, such as differential gene expressionanalysis applications.

[0008] The subject invention provides methods for producing at least oneaRNA molecule corresponding to an mRNA molecule by the following steps:(a) contacting the mRNA molecule with tagged antisense molecule toproduce hybrid structure, where the tagged antisense molecule includesan antisense domain complementary to at least about 20 nt of the mRNAmolecule; and (b) transcribing the aRNA from the hybrid structure or atemplate derivative thereof to produce the at least one aRNA moleculecorresponding to the mRNA molecule. In certain embodiments, the hybridstructure produced in step (a) is converted to a template prior to thetranscription step (b). In certain embodiments, a plurality of aRNAmolecules corresponding to the mRNA molecule is produced. In certainembodiments, the antisense domain is complementary to at least about 25nt, or 30nt, or 50 nt of the mRNA molecule. In certain embodiments, thetagged antisense molecule includes a tag domain that includes an RNApolymerase promoter. In certain embodiments, the mRNA molecule ispresent in a complex nucleic acid mixture. In certain embodiments, themethod further includes separating the hybrid structure from any singlestranded tagged antisense molecules prior to the transcribing step. Incertain embodiments, the method is a method of producing at least oneaRNA molecule for at least two different mRNA molecules, e.g., aplurality of different mRNA molecules, in a sample.

[0009] Also provided are array-based hybridization assays that includethe steps of: (a) generating a population of target nucleic acids usinga population of tagged antisense molecules according to the providedmethods; (b) contacting an array of probe nucleic acids on a surface ofa solid support with the population of target nucleic acids; and (c)detecting hybridization complexes on a surface of the array. In certainembodiments, at least a subset of the probe nucleic acids present on thearray are represented in the population of tagged antisense molecules,and in certain embodiments, all of the probe nucleic acids present onthe array are represented in the population of tagged antisensemolecules.

[0010] Also provided is a set of tagged antisense molecules each for usein preparation of target nucleic acids for hybridization to an array ofprobe nucleic acids corresponding to a plurality of different genes,wherein the set includes at least 20 distinct tagged antisensemolecules, wherein each of the tagged antisense molecules includes anmRNA complementary domain of at least about 20 nt in length, and atleast a subset of said probe nucleic acids on the array is representedin the set of tagged antisense molecules. In certain embodiments, eachof the tagged antisense molecules of the set is of known sequence and ispresent in known amount. In certain embodiments, each of the probenucleic acids on said array is represented in the control set of targetnucleic acids. In certain embodiments, the mRNA complementary domain isat least about 25 nt or at least about 30 nt in length.

[0011] Also provided is a kit for use in producing at least one aRNAmolecule, where the kit includes: (a) at least one RNA polymerasepromoter tagged antisense molecule, e.g., at least about 20, 30 or 50 ntin length; and (b) a DNA dependent RNA polymerase, e.g., T7 RNApolymerase. In certain embodiments, the kit includes a population ofdistinct RNA polymerase promoter tagged antisense molecules. In certainembodiments, the kit further includes ribonucleotides. In certainembodiments, the kit further includes an array of probe nucleic acids ona surface of a substrate.

BRIEF DESCRIPTION OF THE DRAWINGS

[0012]FIG. 1 provides schematic representation of a first embodiment ofthe subject methods in which a hybrid structure is used directly in anRNA polymerase mediated transcription step to produce aRNA.

[0013]FIGS. 2a and 2 b provide a schematic representation of a secondembodiment of the subject methods in which a hybrid structure is firstconverted to a template structure, which template structure is thenemployed in a polymerase mediate transcription step to produce aRNA.

[0014]FIG. 3. Gene expression analysis in rat liver revealed byhybridization of ³²P-labeled aRNA target with Atlas Rat Plastic 4KMicroarray. Set of 1090 of tagged antisense oligonucleotides washybridized with 10 μg of rat liver total RNA and fraction of hybridizedoligonucleotides was amplified, converted to ³²P-labeled aRNA andhybridized with the microarray as described in more details in Example1.

[0015]FIG. 4 provides Table 1, which is a list of the sequences of 1090tagged antisense oligonucleotides. All oligos are single-strandedoxyribooligonucleotides (80-mers) with the same 8 nucleotide tagsequences on the 5′- and 3′-ends. The tag sequences at the 3′- and5′-ends are different from each other. Each antisense oligo correspondindividual rat gene, the gene names listed in a separate column.

[0016]FIGS. 5a and b provides a diagram showing method of mixingantisense pools with different point in printing, and subsequentcomparisons.

[0017]FIG. 6 provides a pictures of a human glass array hybridized withan antisense oligo pool.

DETIALED DISCRIPTION OF THE INVENTION

[0018] Methods for producing a population of distinct aRNA moleculesfrom an initial population of distinct mRNA molecules are provided. Inthe subject methods, an initial mRNA sample is contacted with apopulation of distinct tagged antisense nucleic acid molecules toproduce a population of hybrid molecules. The resultant population ofhybrid molecules, or derivatives thereof (e.g., template structuresproduced there from) is then contacted with a DNA dependent RNApolymerase to produce the population of distinct aRNA molecules via RNApolymerase mediated transcription. Also provided are kits for practicingthese methods. The subject methods find use a variety of differentapplications in which the preparation of aRNA is desired, e.g., thepreparation of nucleic acid targets for use in array based hybridizationapplications.

[0019] Before the subject invention is described further, it is to beunderstood that the invention is not limited to the particularembodiments of the invention described below, as variations of theparticular embodiments may be made and still fall within the scope ofthe appended claims. It is also to be understood that the terminologyemployed is for the purpose of describing particular embodiments, and isnot intended to be limiting. Instead, the scope of the present inventionwill be established by the appended claims.

[0020] In this specification and the appended claims, the singular forms“a,” “an” and “the” include plural reference unless the context clearlydictates otherwise. Unless defined otherwise, all technical andscientific terms used herein have the same meaning as commonlyunderstood to one of ordinary skill in the art to which this inventionbelongs.

[0021] Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range, and any other stated or intervening value in thatstated range, is encompassed within the invention. The upper and lowerlimits of these smaller ranges may independently be included in thesmaller ranges, and are also encompassed within the invention, subjectto any specifically excluded limit in the stated range. Where the statedrange includes one or both of the limits, ranges excluding either orboth of those included limits are also included in the invention.

[0022] Unless defined otherwise, all technical and scientific terms usedherein have the same meaning as commonly understood to one of ordinaryskill in the art to which this invention belongs. Although any methods,devices and materials similar or equivalent to those described hereincan be used in the practice or testing of the invention, the preferredmethods, devices and materials are now described.

[0023] All publications mentioned herein are incorporated herein byreference for the purpose of describing and disclosing those componentsthat are described in the publications which might be used in connectionwith the presently described invention.

[0024] Methods of Producing aRNA

[0025] The subject invention provides methods for producing at least oneaRNA molecule corresponding to an initial mRNA molecule. The subjectinvention can be used to produce aRNA from a single mRNA molecule orsimultaneously produce aRNA from a population of distinct mRNAmolecules. Each of these representative embodiments is described ingreater detail below.

[0026] Production of aRNA From a Single mRNA Molecule

[0027] In certain embodiments, the subject invention provides methods ofproducing one or more, including amplified amounts of, aRNA moleculesfrom an initial mRNA molecule, i.e., a plurality of aRNA moleculescorresponding to the same mRNA molecule. As such, methods of producingamplified amounts of aRNA molecules from an initial mRNA molecule areprovided. By amplified amounts is meant that for each initial mRNAmolecule of interest, multiple corresponding aRNA molecules, where theterm aRNA stands for antisense ribonucleic acid, are produced. Incertain embodiments, the number of corresponding aRNA molecules producedfor each initial mRNA molecule during the subject linear amplificationmethod will be at least about 10, usually at least about 50 and moreusually at least about 100, where the number may be as great as 1000 orgreater.

[0028] By corresponding is meant that the aRNA molecule and the mRNAmolecule have at least a region of the same sequence of ribonucleotides,where the sequence of the entire aRNA molecule produced by the subjectmethods is typically found in the mRNA molecule to which it corresponds.As such, the aRNA molecule has a region that shares a substantial amountof, and typically complete, sequence identity with the sequence of theinitial mRNA molecule to which it corresponds, where substantial amountmeans at least 95%, usually at least 98% and more usually at least 99%,where sequence identity is determined using the BLAST algorithm, asdescribed in Altschul et al. (1990), J. Mol. Biol. 215:403-10 (using thepublished default settings, i.e. parameters w=4 and t=17).

[0029] In practicing the subject methods, the first step is to contact atagged nucleic acid molecule, e.g., tagged DNA molecule, tagged RNAmolecule, and in many embodiments a tagged DNA molecule, with the mRNAmolecule for which the one or more aRNA molecules is to be produced.

[0030] Tagged antisense nucleic acid molecules include at least thefollowing two domains: (1) an mRNA antisense domain complementary to atleast a substantial portion of the mRNA molecule; and (2) at least onetag domain. The mRNA antisense domain is a region that is sufficientlylong to provide for specific hybridization to its corresponding mRNAmolecule and to generate an aRNA molecule of the desired length, asdescribed below. As mentioned above, the mRNA complementary antisenseDNA domain is complementary to a substantial portion of the mRNAmolecule. By “substantial portion” is meant a length of at least about20 nt, usually at least about 50 nt, often at least about 65 nt, 100 ntor longer, including and up to the full length of the mRNA, e.g., 150nt, 200 nt, 300nt or longer. In certain embodiments, the taggedantisense molecules are synthetic nucleic acids, e.g., synthesized byphosphoramidite chemistry, and range in length from about 30 to 150 nt,and often from about 65 to 85 nt.

[0031] The tagged antisense nucleic acids may be polymers of syntheticnucleotide analogs. Such tagged antisense nucleic acids may be preferredin certain embodiments because of their superior stability under assayconditions. Modifications in the native structure, including alterationsin the backbone, sugars or heterocyclic bases, have been shown toincrease intracellular stability and binding affinity. Among usefulchanges in the backbone chemistry are phosphorothioates;phosphorodithioates, where both of the non-bridging oxygens aresubstituted with sulfur; phosphoroamidites; alkyl phosphotriesters andboranophosphates. Achiral phosphate derivatives include3′-O′-5′-S-phosphorothioate, 3′-S-5′-O-phosphorothioate,3′-CH₂-5′-O-phosphonate and 3′-NH-5′-O-phosphoroamidate. Peptide nucleicacids replace the entire ribose phosphodiester backbone with a peptidelinkage. Locked nucleic acids give additional conformational stabilityof sugar moiety due to additional bonds between 2′-carboxil and5′-carboxil or 4′-carboxil groups of deoxyribose. Sugar modificationsare also used to enhance stability and affinity. The α-anomer ofdeoxyribose may be used, where the base is inverted with respect to thenatural β-anomer. The 2′-OH of the ribose sugar may be altered to form2′-O-methyl or 2′-O-allyl sugars, which provides resistance todegradation without comprising affinity. Modification of theheterocyclic bases that find use in the method of the invention arethose capable of appropriate base pairing. Some useful substitutionsinclude deoxyuridine for deoxythymidine; 5-methyl-2′-deoxycytidine and5-bromo-2′-deoxycytidine for deoxycytidine. 5-propynyl-2′-deoxyuridineand 5-propynyl-2′-deoxycytidine have been shown to increase affinity andbiological activity when substituted for deoxythymidine anddeoxycytidine, respectively. Examples of non-naturally occurring basesthat are capable of forming base-pairing relationships include, but arenot limited to, aza and deaza pyrimidine analogues, aza and deaza purineanalogues, and other heterocyclicbase analogues, wherein one or more ofthe carbon and nitrogen atoms of the purine and pyrimidine rings havebeen substituted by heteroatoms, e.g., oxygen, sulfur, selenium,phosphorus, and the like.

[0032] In addition to the antisense domain, the tagged antisense DNAmolecules further include at least one tag domain. The main function ofthe tag domains is to provide a hybrid molecule, as described in greaterdetail below, that be transcribed into aRNA by DNA dependent RNApolymerase mediated transcription or can readily be converted to aderivative template structure that is employed in a such a transcriptionstep. Typically, the tagged antisense DNA molecules include from one totwo tags or tag domains, where the tag domains are typically located atone or both of the termini of the molecule, i.e., the 5′ and 3′ termini.Typically, a population of different tagged antisense DNA moleculesshares the same tag domain or domains.

[0033] The tag domains can be any sequence and typically range in lengthfrom about 4 to 50 nt, sometimes from about 6 to 25 nt and often fromabout 8 to 20 nt. The tag domains are typically not complementary to themRNA sequence opposing them when the antisense tagged DNAs arehybridized to their corresponding mRNA molecules, i.e., the tag domainsdo not hybridize (e.g., under stringent conditions) to their opposingdomains in the mRNA molecule.

[0034] Where the tagged antisense molecules include two different tagdomains, i.e., a domain at each termini, the two different tag domainscould be the same but in certain embodiments they are generallydifferent, i.e., they have a different sequence, where two given tagdomains are considered to have a different sequence if they share lessthan 80% homology, e.g., as determined by BLAST, supra.

[0035] The tag domain(s) can be single or double stranded.

[0036] The tag domain or domains can include a variety of differenttypes of sites and, therefore, functionality. Sites of interest include,but are not limited to: restriction sites/regions/subdomains, primerbinding sites/regions/subdomains, RNA polymerase promotersites/regions/subdomains, etc. Restriction site/region/subdomains ofinterest can include any sequence of nucleotide residues that arerecognized by a restriction endonuclease, e.g., Rsa I, EcoRI, etc.Primer binding sites/regions/subdomains of interest are typically singlestranded and include a primer complementary sequence of at least 6 nt inlength, typically at least 8 nt in length and more typically at least 9nt in length. The primer binding sequences are chosen to hybridize withprimers, typically the 3′ end of primers, where the primers typicallyrange in length from about 10 to 75 nt, usually from about 15 to 35 ntand more usually from about 18 to 25 nt. In certain embodiments, theprimers can include a sequence of an RNA polymerase promoter. Theprimers can find use in primer extension reactions, PCR or otheramplification steps, e.g., in order to amplify tagged antisense DNAmolecules, to convert tagged antisense DNA molecules to template DNAstructures that include the RNA polymerase promoter domain, etc. In yetother embodiments, primers can be employed as a template for antisenseDNA extension reactions, where the extended product can be labeled bydetectable probes. This approach is useful in the generation of labeledhybridization target from non-amplified or amplified antisense DNA.

[0037] In certain embodiments (such as those illustrated in FIG. 1), thetag domain includes includes an RNA polymerase promoter domain. In theseembodiments, the tag domain is a 5′ tag domain that includes the RNApolymerase promoter domain. In these embodiments, the promoter domain islinked in an orientation to permit transcription of the mRNA sensedomain strand or nucleic acid corresponding to the sequence of the mRNAdomain (template strand). A linker oligonucleotide between the promoterand the DNA may be present and, if present, will typically comprisebetween about 5 and 20 bases, but may be smaller or larger as desired.

[0038] The tagged antisense nucleic acid molecule is contacted with themRNA molecule of interest under hybridization conditions sufficient toproduce a duplex structure, i.e., hybrid, made up of the taggedantisense DNA molecule hybridized to the mRNA molecule. In manyembodiments, the two molecules are contacted with each other understringent hybridization conditions in order to produce the desiredduplex or hybrid structure. An example of stringent hybridizationconditions is hybridization at 50° C. or higher and 6.0×SSC (900 mMNaCl/90 mM sodium citrate). Another example of stringent hybridizationconditions is overnight incubation at 42° C. or higher in a solution:50% formamide, 6×SSC (900 mM NaCl, 90 mM trisodium citrate), 50 mMsodium phosphate (pH 7.6), 10% dextran sulfate, and 20 μg/ml denatured,sheared salmon sperm DNA. Stringent hybridization conditions arehybridization conditions that are at least as stringent as the aboverepresentative conditions, where conditions are considered to be atleast as stringent if they are at least about 80% as stringent,typically at least about 90% as stringent as the above specificstringent conditions. Other stringent hybridization conditions are knownin the art and may also be employed.

[0039] Following production of the tagged antisense DNA/mRNA structuresby the above hybridization step, the resultant hybrid/duplex structures,or derivates thereof (such as the template structures described ingreater detail below) aRNA is produced via DNA dependent RNA polymerasemediated transcription. In other words, following production of thetagged nucleic acid/mRNA hybrid or duplex structure, one or more aRNAmolecules is produced from the hybrid or duplex structure or aderivative thereof.

[0040] In certain embodiments, the above described hybrid or duplexstructure is employed directly in aRNA production via RNA polymerasemediated transcription, as depicted in FIG. 1. In these embodiments, thehybrid structure includes a suitable RNA polymerase promoter domain.

[0041] With respect to the RNA polymerase promoter domain that isemployed in the transcription steps of the subject invention, a numberof RNA polymerase promoters may be used. Suitable promoter regions arepromoters that are capable of initiating transcription of an operablylinked DNA sequence, e.g., the anstisense DNA domain of a hybridstructure, in the presence of ribonucleotides and an RNA polymeraseunder suitable conditions.

[0042] The promoter region usually includes between about 15 and 150nucleotides, preferably between about 15 and 25 nucleotides, from anaturally occurring RNA polymerase promoter or a consensus promoterregion, as described in Alberts et al., in Molecular Biology of theCell, 2d Ed., Garland, N.Y. (1989). In general, prokaryotic promotersare preferred over eukaryotic promoters, and phage or virus promotersmost preferred. As used herein, the term “operably linked” refers to afunctional linkage between the affecting sequence (typically a promoter)and the controlled sequence. The promoter regions that find use areregions where RNA polymerase binds tightly to the DNA and contain thestart site and signal for RNA synthesis to begin. In E. coli, typicallythe RNA polymerase molecule covers about 60 nucleotides when it binds tothe DNA. Native strong promoters typically contain two highly conservedDNA sequences, each about six nucleotides long, which are locatedupstream from the start site and separated from each other by about 17nucleotides of unrecognized DNA. A wide variety of promoters are known.Representative promoter regions of interest that find use include SP6,T3 and T7 as described in Chamberlin and Ryan, The Enzymes (ed P. Boyer,Academic Press, New York) (1982) pp 87-108. See also, Enzymology Primerfor Recombinant DNA Technology (Academic Press, 1996).

[0043] In certain embodiments, following hybrid structure production viahybridization, as described above, and prior to transcription, thederivative of the hybrid structure is produced from the hybridstructure. More specifically, the hybrid structure is converted to atemplate structure, which template structure is then employed in thetranscription step. This conversion to template structure can includeone or more different steps, depending on the nature of the initialhybrid structure from which the template structure is to be derived. Incertain embodiments where an RNA polymerase promoter sequence is notpresent in the tag domain, it is necessary to incorporate an RNApolymerase promoter domain into the hybrid structure, as describedabove. Incorporation of the RNA polymerase promoter domain can beaccomplished using any convenient protocol, where representativeprotocols include, but are not limited to: primer extension, fusion PCR,ligation, site-specific recombination, etc.

[0044] In certain embodiments, the mRNA strand of the hybrid structureis converted to DNA, where any convenient protocol for converting mRNAto DNA may be employed. For example, the mRNA strand can be converted toDNA using second strand cDNA synthesis protocols, such as using acombination of RNase H and an enzyme with DNA polymerase activity, e.g.,DNA polymerase 1. Alternatively, a primer extension reaction using aprimer complementary to a 3′ tag domain can be employed to generate asecond strand DNA and therefore a template structure. In yet analternative embodiment, a PCR protocol can be used to generate templatestructure from the initial hybrid structure, e.g., by employing PCRprimers to both the 5′ and 3′ tags and amplifying a template structurefrom the initial hybrid structure.

[0045]FIGS. 2A and B provide a flow diagram of a protocol that includesa step in which the hybrid structure is converted to a templatestructure prior to transcription, as described above.

[0046] As summarized above, following hybrid production and, optionallytemplate structure derivatization of the hybrid, the hybrid or templatestructure derivative thereof is employed in DNA dependent RNA polymerasemediated transcription. In many embodiments, prior to the transcriptionstep, the hybrid structures and/or derivatives thereof, are separatedfrom unhybridized tagged antisense nucleic acids. This separation stepmay be accomplished using any convenient protocol, e.g., via physical(e.g. size separation) and/or chemical/enzymatic (e.g., nuclease)protocols.

[0047] For the transcription step, the presence of the RNA polymerasepromoter region of the hybrid structure or template derivative thereofis exploited for the production of aRNA. To synthesize the aRNA, thehybrid structure or template derivative thereof is contacted with theappropriate RNA polymerase in the presence of the four ribonucleotides,e.g. rGTP, rCTP, rATP and rUTP, under conditions sufficient for RNAtranscription to occur, where the particular polymerase employed will bechosen based on the promoter region present in the ds DNA, e.g. T7 RNApolymerase, T3 or SP6 RNA polymerases, E. coli RNA polymerases, and thelike. Suitable conditions for RNA transcription using RNA polymerasesare known in the art, see e.g. the references described in the RelevantLiterature section, supra. In certain embodiments, the ribonucleic acidstrand of the duplex is separated from the DNA strand prior totranscription, e.g., by enzymatic digestion, chemical modification,dissociation and separation, etc.

[0048] The above two steps result in the production of at least one aRNAmolecule that corresponds to the initial mRNA molecule as describedabove.

[0049] Methods of Producing aRNA From a Plurality of Distinct mRNAMolecules

[0050] As indicated above, in many embodiments, the subject methods areemployed to simultaneously produce one or more, e.g., a plurality orpopulation of, aRNA molecules from a plurality of initial distinct mRNAmolecules, e.g., as is found in an initial mRNA population isolated froma cellular source. In other words, the subject methods of thisembodiment are methods of producing a population of distinct aRNAmolecules that correspond to a population of distinct mRNA molecules. Inother words, one or more aRNA molecules are produced simultaneously froma plurality of distinct initial mRNA molecules (where distinct meansthat the molecules have a different sequence). More specifically, incertain embodiments, the subject methods simultaneously produce one ormore aRNA molecules, e.g., amplified amounts of aRNA molecules, from aplurality of different mRNA molecules. For example, the subject methodscan take an initial population of 10 different mRNA molecules andsimultaneously produce 10 or more aRNA molecules for each of thedistinct mRNA molecules in the initial population.

[0051] The resultant population of aRNA molecules reflects or is arepresentation of the initial mRNA population. In other words, theplurality of distinct aRNA molecules generated from the initial mRNApopulation in similar in terms of copy number to the plurality ofdistinct mRNA molecules in the initial mRNA population. Morespecifically, in certain embodiments, the subject methods simultaneouslyproduce amplified amounts of aRNA, where at least 90% of the distinctmolecules have less than 10 fold, often less than 5 fold and sometimesless than 3-fold differences in copy number from the corresponding copynumber of their corresponding mRNAs in the initial mRNA sample. Theinitial mRNA population that is employed in these embodiments may bepresent in a variety of different samples, where the sample willtypically be derived from a physiological source. The physiologicalsource may be derived from a variety of eukaryotic or prokaryoticsources, with physiological sources of interest including sourcesderived from single cell organisms such as yeast or bacteria andmulticellular organisms, including plants and animals, particularlymammals, where the physiological sources from multicellular organismsmay be derived from particular organs, biological fluids (e.g., blood)or tissues of the multicellular organism, or from isolated cells derivedthere from.

[0052] In obtaining the sample of RNAs to be analyzed from thephysiological source from which it is derived, the physiological sourcemay be subjected to a number of different processing steps, where suchprocessing steps might include tissue/cell homogenation, cell isolation,cell fractionation and cytoplasmic extraction, nucleic acid extractionand the like, where such processing steps are known to the those ofskill in the art. Methods of isolating total or polyA+ RNA from cells,tissues, organs, biological fluids or whole organisms are known to thoseof skill in the art and are described in Maniatis et al., MolecularCloning: A Laboratory Manual (Cold Spring Harbor Press) (1989).Alternatively, at least some of the initial steps of the subject methodmay be performed in situ, as described in U.S. Pat. No. 5,514,545, thedisclosure of which is herein incorporated by reference. In certainembodiments, total RNA comprising mRNA is used for hybridization withtagged antisense nucleic acids molecule in hybrid structure production.

[0053] Following provision of the initial mRNA population, e.g., samplecontaining the initial mRNA population, the initial population iscontacted with a population or set of distinct tagged antisense nucleicacid molecules, where the population or set includes a tagged antisensemolecule for each different mRNA molecule of interest that is present inthe initial population.

[0054] The tagged antisense DNA molecules of the subject sets aredeoxyribonucleic acids, preferably single-stranded full-length orfragments of sequences that hybridize to the corresponding mRNAs. Incertain embodiments, the tagged antisense molecules are synthetic singlestranded deoxyribonucleotides complimentary to gene specific portions ofthe mRNAs of interest. In certain embodiments, all of the taggedantisense nucleic acids have the same size (e.g., the are all 80 mers)and have the same or at least similar sized mRNA complementary region ordomain, which typically ranges from about 25 to 70 nt and often fromabout 45 to 60 nt. Furthermore, in certain embodiments the taggedantisense nucleic acids have similar GC content, which often ranges fromabout 40 to 80%, usually from about 45 to 60%, and similar meltingtemperatures (where there is typically not more than a 10° C. variationin this parameter and often not more than a 5° C. variation in thistemperature). Among the population of tagged antisense molecules, all ofthe tag domains are constant or same in many embodiments, with the onlyvariation occurring in the mRNA complementary regions of the taggedantisense nucleic acids.

[0055] A feature of the sets of target nucleic acids is that theyinclude at least one tagged antisense molecule that is complementary toeach mRNA of a predetermined collection or set of mRNAs of interest,e.g., a collection or set of mRNAs that is represented by probe nucleicacids on a nucleic acid microarray. In other words, each of the distinctmRNAs of a given predetermined set or collection of distinct mRNAs arerepresented in the set of tagged antisense nucleic acid molecules. Forexample, where a given predetermined set of mRNAs of interest includes500 distinct mRNAs which are distinct from each other based on sequence,a set of tagged antisense nucleic acid molecules includes at least 500different tagged antisense nucleic acid molecules—one for each probenucleic acid on the array.

[0056] The number of unique tagged antisense molecules (where any twosequences are unique if they differ from each other in terms ofsequence, where the difference may be a minimal as a 1 to 10 basedifference) in the set or pool of tagged mRNA antisense molecules will,in most embodiments, be at least about 10, 20, 50, 100, 200 or morewhere the number may be as high as about 1,000; 20,000 or higher, but inmany embodiments will not exceed about 10,000; 5,000, or 1,000.

[0057] In certain embodiments, the sets include a tagged antisensenucleic acid for each mRNA that may be present in the sample. In otherwords, the set includes a tagged antisense nucleic acid for eachdifferent mRNA molecule that may be present in the sample.

[0058] In yet other embodiments, the sets are sets that include arepresentative or representational number of tagged antisense molecules.As the subject sets include a representational number of taggedantisense molecules, the total number of different tagged mRNA antisensemolecules in any given set will be only a fraction of the total numberof different or distinct mRNAs in the sample that is employed togenerate the aRNA, where the total number of tagged antisense moleculesin the set will generally not exceed 80%, usually will not exceed 60-50%and more usually will not 40-20% of the total number of distinct mRNAsin the original sample, e.g., the total number of distinct messengerRNAs (mRNAs) in the original sample. Any two given RNAs in a sample willbe considered distinct or different if they comprise a stretch of atleast 100 nucleotides in length in which the sequence similarity is lessthen 95% or lower, as determined using the FASTA program (defaultsettings). As the sets of tagged antisense molecules comprise only arepresentational number of target nucleic acids compared to the initialmRNA population of the sample from which the aRNA population is to beproduced, with sources comprising from 5,000 to 50,000 distinct mRNAs,the number of different tagged antisense molecules in the set typicallyranges from about 20 to 40,000 or 20 to 10,000, usually from about 50 to2,000 or 50 to 30,000 and more usually from about 100 to 20,000 andsometimes from about 75 to 1500.

[0059] In some embodiments, a feature of the sets of tagged antisensemolecules is that the concentration of each tagged antisense moleculepresent in the set is known. In other words, the amount of eachindividual RNA polymerase promoter tagged antisense DNA molecule in thecontrol set is known. For example, an equal weight amount of eachdistinct tagged antisense molecules may present in the mixture. Incertain embodiments, an equal molar amount or equimolar amount of eachtagged antisense molecule may be present. In yet other embodiments,different known amounts or ratios of the various constituent taggedantisense molecules may be present. However, in any set of taggedantisense molecules employed according to the subject invention, theamount of each constituent member present in the set is known, either inabsolute terms or in terms relative to each other.

[0060] The sets of tagged antisense molecules are further characterizedin that at least two different gene functional classes are typicallyrepresented in a given set, where the number of different functionalclasses of genes represented in the a given set will generally be atleast 3, and will usually be at least 5. In other words, the sets oftagged antisense molecules comprise nucleotide sequences complementaryto RNA transcripts of at least 2 gene functional classes, usually atleast 3 gene functional classes, and more usually at least 5 genefunctional classes. Gene functional classes of interest includeoncogenes; genes encoding tumor suppressors; genes encoding cell cycleregulators; stress response genes; genes encoding ion channel proteins;genes encoding transport proteins; genes encoding intracellular signaltransduction modulator and effector factors; apoptosis related genes;DNA synthesis/recombination/repair genes; genes encoding transcriptionfactors; genes encoding DNA-binding proteins; genes encoding receptors,including receptors for growth factors, chemokines, interleukins,interferons, hormones, neurotransmitters, cell surface antigens, celladhesion molecules etc.; genes encoding cell-cell communicationproteins, such as growth factors, cytokines, chemokines, interleukins,interferons, hormones etc.; and the like.

[0061] Of particular interest are sets of tagged antisense molecules inwhich each of the genes collectively listed in the tables of thefollowing applications are represented in the control set: U.S. patentapplication Ser. No. 08/859,998; U.S. patent application Ser. No.08/974,298; U.S. patent application Ser. No. 09/225,998; U.S.application Ser. No. 09/221,480; U.S. application Ser. No. 09/222,432;U.S. application Ser. No. 09/222,436; U.S. application Ser. No.09/222,437; U.S. application Ser. No. 09/222,251; U.S. application Ser.No. 09/221,481; U.S. application Ser. No. 09/222,256; U.S. applicationSer. No. 09/222,248; and U.S. application Ser. No. 09/222,253; thedisclosures of which are incorporated herein by reference.

[0062] Another feature of the sets of tagged antisense molecules is thatthey are synthetic nucleic acids and not isolated from a biologicalsource. The sets of tagged antisense molecules may be generated usingany convenient protocol.

[0063] As described above, the initial mRNA population of distinctmRNAs, which includes the mRNAs of interest if present, is contactedwith the population or set of tagged antisense molecules underconditions sufficient to produce hybrid or duplex structures betweencomplementary nucleic acids, e.g., hybridization conditions, typicallystringent hybridization conditions. In this step duplex or hybridstructures are produced between any tagged antisense molecules presentin the employed set that have a complementary mRNA strand in the sample.In many embodiments, the hybridization conditions employed are thosethat maintain the integrity of the mRNA and tagged antisense molecules,like RNase-free conditions. A representative example of such conditionsis: 50° C. in (5× SSPE, pH 7.5, 6M urea, 1×SUPERase·in™ RNase inhibitor(Ambion, Inc.) overnight.

[0064] Following production of the hybrid or duplex structures, in manyembodiments any remaining single stranded or unbound tagged antisensemolecules that are present in the mixture, i.e., those tagged antisensemolecules that do not have a complementary mRNA molecule present in theinitial mRNA sample, are separated from the resultant duplex structures.Separation may be accomplished using any convenient protocol (where anumber of different protocols are known in the art), including enzymaticor chemical modification and physical separation protocols, etc. Inenzymatic protocols, a nuclease that selectively degrades or modifiessingle stranded and not double stranded nucleic acids, and at leastsingle stranded DNAs, may be employed, where representative singlestrand specific nucleases include, but are not limited to: S1 nuclease,mung bean nuclease, ribonuclease A, T1, and the like. Physicalseparation protocols of interest include, but are not limited to: gelelectrophoresis, chromatography, precipitation, centrifugation,filtration, binding to immobilized ligands specific for the singledstranded tagged antisense molecules or a region thereof, etc. Inchemical modification, the non-bound antisense tagged nucleic acidmolecules, but not the hybrid structures, are selective modified(inactivated) by a chemical reactant, such as glyoxal,dimehtylsulfoxide, etc.

[0065] Following production of the hybrid structures, the hybridstructures are employed directly in aRNA transcription or, are convertedto template derivatives which are then employed in aRNA transcription.Where the hybrid structures are converted to template structures priorto aRNA transcription, any of the protocols discussed above forproduction of template derivatives from hybrid structures may beemployed.

[0066] Next, aRNA is transcribed from each of the hybrids or templatederivatives thereof as described above. For this transcription step, theRNA polymerase promoter region of the hybrid or template structures isexploited for the production of aRNA. To synthesize the aRNA, thehybrids or templates are contacted with the appropriate RNA polymerasein the presence of the four ribonucleotides, e.g. rGTP, rCTP, rATP andrUTP, under conditions sufficient for RNA transcription to occur, wherethe particular polymerase employed will be chosen based on the promoterregion present in the ds DNA, e.g. T7 RNA polymerase, T3 or SP6 RNApolymerases, E. coli RNA polymerases, and the like. Modifiednucleotides, e.g., fluoro rNTP (such as Cy3-rUTP) biotin-rUTP,allylamine rNTP, etc., could be used in order to provide higherstability, hybridization efficiency, post synthesis labeling, etc., ofthe synthesized aRNA. Suitable conditions for RNA transcription usingRNA polymerases are known in the art, see e.g. the references describedin the Relevant Literature section, supra.

[0067] The resultant aRNA produced by the subject methods finds use in avariety of applications. For example, the resultant aRNA can be usedfor: (1) cDNA library construction; as target or for use in generationof target nucleic acids for use with microarrays, e.g., in expressionprofiling analysis; construction of “driver” for subtractivehybridization assays; and the like.

[0068] For example, the aRNA produced by the subject invention finds usein studies of gene expression in mammalian cell or other cellpopulations. The cells may be individual cells or tissue derived cells,e.g., tissue derived from a solid organs, such as brain, spleen, bone,heart, vascular, lung, kidney, liver, pituitary, endocrine glands, lymphnode, dispersed primary cells, tumor cells, or the like. In theserepresentative methods, one typically identifies nucleic acid sequencesthat vary in abundance among different populations, such as in comparingmRNA expression among different tissues or within the same tissueaccording to physiologic state known as subtractive hybridizationassays.

[0069] Depending on the particular intended use of the subject aRNA, theaRNA may be labeled. A variety of different labels may be employed,where such labels include fluorescent labels, isotopic labels, enzymaticlabels, particulate labels, etc. For example, suitable labels includefluorochromes, e.g. fluorescein isothiocyanate (FITC), rhodamine, TexasRed, phycoerythrin, allophycocyanin, 6-carboxyfluorescein (6-FAM),2′,7′-dimethoxy-4′,5′-dichloro-6-carboxyfluorescein (JOE),6-carboxy-X-rhodamine (ROX),6-carboxy-2′,4′,7′,4,7-hexachlorofluorescein (HEX), 5-carboxyfluorescein(5-FAM) or N,N,N′,N′-tetramethyl-6-carboxyrhodamine (TAMRA), cyaninedyes, e.g. Cy5, Cy3, BODIPY dyes, e.g. BODIPY 630/650, Alexa542, etc.Suitable isotopic labels include radioactive labels, e.g. ³²P, ³³P, ³⁵S,³H. Other suitable labels include size particles that possess lightscattering, fluorescent properties or contain entrapped multiplefluorophores. The label may be a two stage system, where the target DNAis conjugated to biotin, haptens, etc. having a high affinity bindingpartner, e.g. avidin, specific antibodies, etc. The binding partner isconjugated to a detectable label, e.g. an enzymatic label capable ofconverting a substrate to a chromogenic product, a fluorescent label,and isotopic label, etc.

[0070] Differential Gene Expression Assays

[0071] As indicated above, one application in which the aRNA moleculesproduced by the subject methods find use is as target nucleic acids indifferential gene expression analysis. Using the subject methods, onecan produce labeled aRNA molecules, or labeled derivatives of suchmolecules, e.g., first or second strand cDNA molecules, generated fromthe aRNA molecules, etc., which are then used as target nucleic acids indifferential gene expression analysis. In certain embodiments, thelabeled target nucleic acids produced according to the subject inventionrepresent the entire or whole mRNA profile of the sample being assayed.In other embodiments, the labeled target nucleic acids producedaccording to the subject invention provide a representation of the totalRNA profile of the particular source from which the labeled nucleicacids are generated, since not all of the mRNAs present in the initialsample are represented in the set of tagged antisense molecules employedto generate the mRNA. Accordingly, the labeled nucleic acids find use incomparing the characteristic RNA profiles of different physiologicalsources and identifying differences in the RNA profiles betweendifferent physiological source. Comparison of the RNA profiles of two ormore physiological sources finds particular use in methods ofidentifying differential gene expression in two physiological samples,such as cells or tissues derived from the same or different individualorganisms, where the tissues may represent different diseased or normalstates, different organ or tissue types, etc.

[0072] The labeled nucleic acids of the plurality of physiologicalsources may be compared in a number of different ways. Thus, one maycompare the labeled nucleic acids from each source by separatelyresolving the labeled nucleic acids from each source under substantiallyidentical electrophoretic conditions to yield an electrophoretic patternof resolved bands for each of the different populations of labelednucleic acids. The resultant electrophoretic patterns can then beresolved to identify differences between the labeled nucleic acidpopulations, which differences can then be attributed to differences inthe RNA profiles of the each of the physiological sources, where suchdifferences can, in turn, be attributed to difference in geneexpression. See Liang & Pardee, Science (1992) 257: 967. Conveniently,electrophoretic separation under identical electrophoretic conditionscan be achieved by running the labeled nucleic acids derived from eachphysiological source of interest in separate, side by side lanes on aslab gel. Automated electrophoretic machines as described in U.S. Pat.Nos. 5,410,412; 5,275,710; 5,217,591; and 5,104,512, the disclosures ofwhich are herein incorporated by references, may be employed to resolvethe labeled nucleic acids. In a modification of the above, where eachset of labeled nucleic acids or targets of each physiological source hasbeen labeled with a distinct and distinguishable label, the opportunityarises to resolve the nucleic acids in the same electrophoretic medium,e.g. the same column or in the same lane of a slab, thereby ensuringthat the nucleic acids are resolved under identical electrophoreticconditions.

[0073] Alternatively, one may hybridize the labeled nucleic acids topredefined arrays of probe polymeric molecules stably associated withthe surface of a substrate, where the probe polymeric molecules arecapable of sequence specific base pair hybridization with complementarylabeled target nucleic acids. A variety of different arrays which may beused are known in the art. The polymeric or probe molecules of thearrays may be nucleic acids, e.g., oligonucleotides/polynucleotides, orhybridizing analogues or mimetics thereof, including: nucleic acids inwhich the phosphodiester linkage has been replaced with a substitutelinkage, such as phophorothioate, methylimino, methylphosphonate,phosphoramidate, guanidine and the like; nucleic acids in which theribose subunit has been substituted, e.g. hexose phosphodiester; peptidenucleic acids; and the like. In many embodiments, the length of theprobes ranges from 10 to 1000 nts, where oligonucleotide probes usuallyrange from 15 to 150 nts and more usually from 25 to 100 nts in length,and polynucleotide probes usually range in length from 150 to 1000 nts,where the polynucleotide probes may be single or double stranded,usually single stranded, and may be PCR fragments amplified from cDNA.

[0074] The probe molecules on the surface of the substrates in certainembodiments correspond to the set of tagged antisense molecules employedto generate the set of aRNA used as, or to generate, the labeled targetnucleic acids. The term “correspond” is used in this instance to meanthat all of the distinct tagged antisense molecules present in the setused to generate the aRNA have a corresponding probe on the array, wherecorrespond means that the probe and the tagged antisense molecule hasthe same (at least 95% homology) sequence for the mRNA binding domain,they hybridize to the same mRNA molecule under stringent conditions.

[0075] The substrates with which the probe molecules are stablyassociated may be fabricated from a variety of materials, includingplastics, ceramics, metals, gels, membranes, glasses, and the like.

[0076] A variety of different methodologies have been developed forproducing arrays of probes stably associated to the surface of asubstrate. Representative methodologies include spotting methods, inwhich probes are immobilized or spotted on the surface of substrates asdescribed in WO 95/35505 the disclosure of which is herein incorporatedby reference, and methods in which the probes are synthesized or grownon the surface of the substrates, such as EP 0 373 203 B1 and U.S. Pat.No. 5,445,934, the disclosures of which are herein incorporated byreference. Arrays of probes spotted onto nylon membranes are describedin Lennon & Lerach, Trends in Genetics (1991) 7:314-317; Gress et al.,Mammalian Genome (1992) 3:609-619; Meier-Ewert et al., Nature (1993)361:375-376; Nguyen et al., Genomics (1995) 29:207-216; Zhao et al.,Gene (1995) 156:207-213; Takahashi et al., Gene (1995) 164:219-217;Milosavlijevic et al., Genome Research (1996) 6:132-141; Pietu et al.,Genome Research (1996) 6:492-503; and Drmanac, Science (1993)260:1649-1652. Arrays of probes spotted onto the surface of modifiedmicroscope glass slides are described in Shena et al., Science(1995)270: 467470 and Shalon et al., Genome Research (1996) 6: 639-645.Arrays in which the probes have been grown on the surface of a substrateare described in Lockhart et al., Nature Biotechnology (1996) 14:1675.

[0077] Of particular interest for use in the analysis of differentialgene expression in various human, mouse and rat physiological sourcesare the arrays sold under the trademark Atlas™ by Clontech Laboratories,Inc., which product line includes nylon, glass and plastic arrays ofprobe nucleic acids.

[0078] In these hybridization assays, the target nucleic acid iscontacted with the array under hybridization conditions, where suchconditions can be adjusted, as desired, to provide for an optimum levelof specificity in view of the particular assay being performed. Suitablehybridization conditions are well known to those of skill in the art andreviewed in Maniatis et al, supra and WO 95/21944. Of particularinterest in many embodiments is the use of stringent conditions duringhybridization, i.e. conditions that are optimal in terms of rate, yieldand stability for specific probe-target hybridization and provide for aminimum of non-specific probe/target interaction. Stringent conditionsare known to those of skill in the art. In the present invention,stringent conditions are typically characterized by temperatures rangingfrom 15° C. to 35° C., usually 20° C. to 30° C. less than the meltingtemperature of the probe target duplexes, which melting temperature isdependent on a number of parameters, e.g. temperature, buffercompositions, size of probes and targets, concentration of probes andtargets, etc. As such, the temperature of hybridization typically rangesfrom about 55° C. to 70° C., usually from about 60° C. to 68° C. In thepresence of denaturing agents, the temperature may range from about 35°C. to 45° C., usually from about 37° C. to 42° C. The stringenthybridization conditions are further typically characterized by thepresence of a hybridization buffer, where the buffer is characterized byone or more of the following characteristics: (a) having a high saltconcentration, e.g. 3 to 6×SSC (or other salts with similarconcentrations); (b) the presence of detergents, like SDS (from 0.1 to20%), triton X100 (from 0.01 to 1%), Monidet NP40 (from 0.1 to 5%) etc.;(c) other additives, like EDTA (typically from 0.1 to 5 mM),tetramethylammonium chloride; (d) accelerating agents, e.g. PEG, dextransulfate (5 to 10%), CTAB, SDS and the like; (e) denaturing agents, e.g.formamide, urea etc.; and the like.

[0079] In analyzing the differences in the population of labeled targetnucleic acids generated from two or more physiological sources using thearrays described above, in certain embodiments each population oflabeled target nucleic acids are separately contacted to identical probearrays or together to the same array under conditions of hybridization,preferably under stringent hybridization conditions, such that labeledtarget nucleic acids hybridize to complementary probes on the substratesurface. In yet other embodiments, labeled target nucleic acids arecombined with a set of distinguishably labeled standard or controltarget nucleic acids followed by hybridization of the combinedpopulations to the array surface, as described in application Ser. No.09/298,361; the disclosure of which is herein incorporated by reference.

[0080] In certain preferred embodiments, the labeled target nucleicacids generated according to the present methods is used in conjunctionwith control nucleic acids as described in application Ser. Nos.09/298,361 and 09/750,452, the disclosures of which are hereinincorporated by reference, as well as in international applicationserial no. PCT/US00/10894 published as WO 00/65095.

[0081] In these embodiments, the tagged antisense nucleic acids arestructurally as similar as possible to the control target nucleic acidsthat are employed in the assay, e.g., both sets of tagged antisensenucleic acids and control nucleic acids are oligonucleotides with thesame or similar sequences. In other words, the structure of the controltarget nucleic acids should be similar to that of the aRNA in order tomaximally imitate the hybridization of the aRNA with which they are tobe employed.

[0082] Where all of the target sequences comprise the same label,different arrays will be employed for each physiological source (wheredifferent could include using the same array at different times).Alternatively, where the labels of the targets are different anddistinguishable for each of the different physiological sources beingassayed, the opportunity arises to use the same array at the same timefor each of the different target populations. Examples ofdistinguishable labels are well known in the art and include: two ormore different emission wavelength fluorescent dyes, like Cy3 and Cy5,two or more isotopes with different energy of emission, like ³²P and³³P, gold or silver particles with different scattering spectra, quantumdot particles, labels which generate signals under different treatmentconditions, like temperature, pH, treatment by additional chemicalagents, etc., or generate signals at different time points aftertreatment. Using one or more enzymes for signal generation allows forthe use of an even greater variety of distinguishable labels, based ondifferent substrate specificity of enzymes (alkalinephosphatase/peroxidase).

[0083] Following hybridization, non-hybridized labeled nucleic acid isremoved from the support surface, conveniently by washing, generating apattern of hybridized nucleic acid on the substrate surface. A varietyof wash solutions are known to those of skill in the art and may beused.

[0084] The resultant hybridization patterns of labeled nucleic acids maybe visualized or detected in a variety of ways, with the particularmanner of detection being chosen based on the particular label of thetarget nucleic acid, where representative detection means includescintillation counting, autoradiography, fluorescence measurement,calorimetric measurement, light emission measurement, light scattering,and the like.

[0085] Following detection or visualization, the hybridization patternsmay be compared to identify differences between the patterns. Wherearrays in which each of the different probes corresponds to a known geneare employed, any discrepancies can be related to a differentialexpression of a particular gene in the physiological sources beingcompared.

[0086] The provision of appropriate controls on the arrays permits amore detailed analysis that controls for variations in hybridizationconditions, cross-hybridization, non-specific binding and the like.Thus, for example, in a preferred embodiment, the hybridization array isprovided with normalization controls. These normalization controls areprobes complementary to control target sequences added in a knownconcentration to the sample. Where the overall hybridization conditionsare poor, the normalization controls will show a smaller signalreflecting reduced hybridization. Conversely, where hybridizationconditions are good, the normalization controls will provide a highersignal reflecting the improved hybridization. Normalization of thesignal derived from other probes in the array to the normalizationcontrols thus provides a control for variations in hybridizationconditions. Normalization control is also useful to adjust (e.g.correct) for differences which arise from the array quality, the mRNAsample quality, efficiency of first-strand synthesis, etc. Typically,normalization is accomplished by dividing the measured signal from theother probes in the array by the average signal produced by thenormalization controls. Normalization may also include correction forvariations due to sample preparation and amplification. Suchnormalization may be accomplished by dividing the measured signal by theaverage signal from the sample preparation/amplification control probes.The resulting values may be multiplied by a constant value to scale theresults.

[0087] In certain embodiments, normalization controls are oftenunnecessary for useful quantification of a hybridization signal. Thus,where optimal probes have been identified, the average hybridizationsignal produced by the selected optimal probes provides a goodquantified measure of the concentration of hybridized nucleic acid.However, normalization controls may still be employed in such methodsfor other purposes, e.g. to account for array quality, mRNA samplequality, etc.

[0088] One may use the subject methods in the differential expressionanalysis of: (a) diseased and normal tissue, e.g. neoplastic and normaltissue, (b) different tissue or tissue types; (c) developmental stage;(d) response to external or internal stimulus; (e) response totreatment; and the like. The subject arrays therefore find use in broadscale expression screening for drug discovery, diagnostics and research,as well as studying the effect of a particular active agent on theexpression pattern of genes in a particular cell, where such informationcan be used to reveal drug toxicity, carcinogenicity, etc.,environmental monitoring, disease research and the like.

[0089] Kits

[0090] Also provided are kits for use in the subject invention, wheresuch kits may include one or more containers, each with one or more ofthe various reagents employed in the subject methods, where the reagentsmay be present in concentrated form. Thus, the kits may include one ormore reagents that are employed to generate aRNA from mRNA according tothe present invention, where the kits typically include at leastsolution with one type of tagged antisense molecule, where in manyembodiments the kits will include a set of a plurality of distincttagged antisense molecules. The kits may also include additionalreagents used in the subject methods, including: buffers, theappropriate nucleotide triphosphates (e.g., rATP, rCTP, rGTP and UTP,including labeled or modified versions thereof, e.g., for generatinglabeled target nucleic acids), RNA polymerase, e.g., T7 RNA polymerase,labeled initiator oligonucleotides used by RNA polymerase to prime aRNAsynthesis, RNase inhibitors, etc. Also included may be purification orisolation reagents, e.g., for isolating or purifying the taggedantisense/mRNA hybrids, where such reagents included chromatographycolumns, binding/washing solutions, modification nucleases, glyoxal, andthe like. Other reagents of interest include PCR primers, PCR reagents,etc. In certain embodiments, e.g., where the kits are specificallydesigned for use in differential gene expression analysis, the kits mayfurther include one or more arrays for use in the gene expressionanalysis application.

[0091] In addition to above mentioned components, the subject kitstypically further include instructions for using the components of thekit to practice the subject methods with the subject devices, e.g., auser manual. The instructions for practicing the subject methods aregenerally recorded on a suitable recording medium. For example, theinstructions may be printed on a substrate, such as paper or plastic,etc. As such, the instructions may be present in the kits as a packageinsert, in the labeling of the container of the kit or componentsthereof (i.e., associated with the packaging or subpackaging) etc. Inother embodiments, the instructions are present as an electronic storagedata file present on a suitable computer readable storage medium, e.g.CD-ROM, diskette, etc. In yet other embodiments, the actual instructionsare not present in the kit, but means for obtaining the instructionsfrom a remote source, e.g. via the internet, are provided. An example ofthis embodiment is a kit that includes a web address where theinstructions can be viewed and/or from which the instructions can bedownloaded. As with the instructions, this means for obtaining theinstructions is recorded on a suitable substrate.

[0092] Commercial kits based on the inventive concept have been shown tobe effective in actual practice, see Dudley, A. M. et al. (2002) Proc.Natl. Acad. Sci. USA 99:7554-7559, incorporated here by reference.

[0093] The following examples are offered by way of illustration and notby way of limitation.

EXPERIMENTAL Example 1

[0094] A. Isolation and Preparation of Test Target Ribonucleic AcidsFrom Rat Tissue Sample.

[0095] As described in greater detail below, the Atlas Plastic Ratmicroarray (BD Clontech, Palo Alto, Calif., Cat. 7909-1) was used toscreen a rat liver tissue sample to determine gene expression of each ofthe 1000 genes in a rat sample. The tissue sample was tested forexpression by isolating total RNA from the sample, hybridizing the totalRNA with mixture of tagged antisense oligonucleotides, converting thehybridized oligonucleotides to template and subsequently transcribingthe template oligonucleotides into labeled aRNA. This labeled pool ofaRNA was then used as test target nucleic acids for hybridizationagainst the probe nucleic acids on the array.

[0096] Total RNA was isolated from homogenized rat liver tissue using anATLASPURE RNA isolation (BD CLONTECH, Palo Alto Calif.) according to themanufacturer's protocols. The total RNA of the tissue sample was used togenerate target aRNA for hybridization to the probe array.

[0097] B. Hybridization of Tagged Antisense Oligonucleotides with TotalRNA.

[0098] At the first step, a mixture of 1,090 rat tagged antisense oligoswas hybridized with rat total liver RNA under the following conditions.The sequences of the individual rat oligos are provided in Table 1appearing in FIG. 4. 10 μg of total liver RNA was mixed with 2 μl of 20uM solution of rat 1,000 tagged antisense oligo set, 2 μl of 4 uMsolution of Bio-r(U)30-Bio (oligo r(U)30 biotinilated at the 5′ and3′-ends) and water to 14 μl of total volume, incubate in thermocycler at75° C. for 3 min then cooled the mix to 50° C. Then 12.5 μl of 2×HWBUbuffer (2M NaCl, 100 mM HEPES-NaOH, pH 7.5, 10 mM EDTA, 8M urea) and 1μl of SUPERASE-IN (Ambion, Austin, Tex.) were added to the mix andcontinue hybridization at 50° C. overnight.

[0099] At the second step the hybridized complexes of RNA and taggedantisense oligos were purified on a streptavidin magnetic beads (Dynal,Oslo, Norway). The hybridization mix was mixed with 20 μl ofstreptavidin magnetic beads (Dynal, Oslo, Norway) prewashed in 1×HWBbuffer and resuspended in 75 μl of 1×HWB buffer (1M NaCl, 50 mMHepes-NaOH, pH 7.5, 5 mM EDTA). Then we continued binding reaction for30 min in the ThermoMixer at 1200 rpm at 50° C., collected particles inMPC-S magnetic stand at the room temperature, wash the beads twice in500 μl of 1×HWB buffer at room temperature, then ones in 500 μl of2×SSPE and ones in 100 μl of 1×RNase H buffer (50 mM HEPES-NaOH, pH 7.5;150 mM NaCl; 4 mM MgCl₂). The fraction of antisense oligos bound to RNAwas eluted by resuspending the particles in 25 μl of RNase H buffercontaining 200 units/ml RNase H and incubating at 25° C. for 30 min.

[0100] C. Generation of Labeled aRNA Target.

[0101] At the first step the fraction of tagged antisense oligoshybridized to RNA was converted to template comprising T7 promoter byPCR. PCR was conducted in 50-μl reaction, containing the followingcomponents: 25 μl of antisense oligos from stage B, 1×Advantage buffer(BD Clontech, Palo Alto, Calif.), 200 μM of each dATP, dGTP, dCTP anddTTP, 1×Advantage enzyme mix and 0.2 μM of each PCR primer T7-adc8F(5′-TMTACCACTCACTATAGGGAGACTCTCACC-3′) and Rev2-17(5′-CGCTCGAGAGGGAGAGT-3′). PCR cycling parameters were as following: 1cycle at 92° C. for 2 min, followed by 2 cycles at 92° C. for 30 sec,45° C. for 15 sec and 68° C. for 15 sec, followed by 21 cycles at 92° C.for 30 sec and 68° C. for 30 sec. PCR product was analyzed by agarosegel electrophoresis and purified by Nucleospin PCR purification kit (BDClontech, Palo Alto, Calif.) and quantitated by A260 nm UV absorbance.

[0102] At the second step the aRNA labeled by detectable label wasgenerated using template comprising T7 promoter at the 5′end ofantisense oligos. ³²P-labeled aRNA was synthesized in 25 μl reactioncontaining: 40 ng of template from first step, 1×MegaScript buffer(Ambion, Austin, Tex.), 500 μM of each rATP, rGTP and rCTP, 25 μCi of³²P-rUTP (400 Ci/mmol) and 2 μl of T7 RNA polymerase (Ambion, Austin,Tex.). The T7 reaction mix was incubated at 37° C. for 30 min andpurified through Nucleospin RNA II purification kit (BD Clontech, PaloAlto, Calif.).

[0103] D. Hybridization and Detection of Target Sequences on Array

[0104] The 100 μl of ³²P-labeled probe generated at stage C wasdenatured at 75° C. for 3 min, mixed with 10 ml of PlasticHyb solution(BD Clontech, Palo Alto, Calif.) and hybridized overnight with Atlas RatPlastic 4K microarray (BD Clontech, Palo Alto, Calif.) in a bottles inhybridization incubator at 60° C. and 10 rpm. After hybridization theplastic films were washed three times by 200 ml of prewarmed at 60° C.wash solution (0.1×SSC, 0.1% SDS) for 15 min each followed by rinse in0.1×SSC. The dried microarray was exposed to phosphorimaging screenovernight and screen was scanned in Phosphorimager Storm at 50 μmresolution (Molecular Dynamics, Mountain View, Calif.). FIG. 3demonstrates pattern of the signals corresponding expressed genesrevealed by labeled aRNA target.

[0105] In an alternative to the above protocol, the aRNA could belabeled by any detectable group (for example fluorescent) using any wellknown in art protocol and signals detected by fluorescence,chemilumenescence, electrochemical, light scattering, surface plasmaresonance, etc. approach. For differential gene expression analysis atleast two aRNA targets could be labeled by the same label, hybridizedwith the same array type and differences in pattern of the signals willreflect the differences in the level of expression of the genes. Inother case two aRNA targets could be labeled by different labels (forexample ³³P and ³²P, Cy3 and Cy5, etc) and hybridized together with thesame array. In this approach the ratio in normalized intensities ofdifferent labels for every dot on the array will reflect the differencesin the level of expression of particular genes. In other preferredapproach the direct comparisons of the amounts of expression on eacharray are carried out by determining the ratio of the intensity of thetest aRNA target hybridization with the intensity of the control target(calibration standard) hybridization. Calibration standards (asdescribed in published PCT application no. WO 00/65095; the priorityapplication of which is herein incorporated by reference) could be theset of labeled aRNA corresponding starting population of taggedantisense oligonucleotides. Once determined, these ratios allow forcorrection of experimental variation between the arrays, and thus allowa direct comparison of the levels of gene expression in the biologicalsamples.

Example 2

[0106] In a specific embodiment of the present invention, the inventorshave developed a research tool product, available to the public. TheseBD Atlas™ Antisense Oligo Mixes provide the ultimate calibration andquality control standard for microarray experiments. Eachspecies-specific mix contains a complete antisense complement to thegenes represented on Clontech BD Atlas™ Glass Microarrays at the time ofmanufacture, providing the most precise and reliable microarray controlavailable. Antisense Oligo Mixes are useful both as a standard to verifylot-to lot consistency in microarray printing and for normalization ofexperimental data to ensure that changes in hybridization intensityreflect genuine shifts in gene expression. Because the mixes aremanufactured on a very large scale, encompassing thousands of kits perbatch, the user can reliably compare results from mixes shipped atdifferent times.

[0107] High-quality array oligos exhibit two main characteristics:efficient hybridization with the intended target and minimalcross-hybridization with other sequences. When these two parameters areconsidered, a stronger array signal (or even a high signal to noiseratio) is not necessarily a better signal. In fact, in many cases astrong signal is the result of cross-hybridization and/or bad oligosynthesis.

[0108] Based upon many years of ordering and printing oligos, theinventors have determined that about 25% of commercially synthesizedoligos are incapable of producing a strong, specific hybridizationsignal. This failure rate is based on a survey of oligos synthesizedfrom four well-established vendors. Failed oligos often show anincreased signal intensity, which is likely due to high non-specificcross hybridization, which may in turn result from inadvertent synthesisof a degenerate oligo.

[0109] Because of the wide variability in oligo synthesis quality, BDBiosciences Clontech has chosen to invest in antisense oligo testing.This analysis ensures that each oligo consists of the intended sequenceand provides a more stringent test than hybridization with “universal”RNA mixtures. Each long oligo included on our pre-made arrays, customarrays, or in Ready-to-Print Long Oligo sets has been antisense testedin two ways. First, Klenow enzyme is used to synthesize the antisensesequences for all oligos on a test array. These antisense oligos arelabeled in groups of 200-300 and hybridized to a BD Atlas microarraycontaining several thousand oligos. Printed (sense) oligos that do noteffectively hybridize to their antisense are resynthesized along withthe corresponding antisense oligo.

[0110] Effective hybridization is defined as production of a signalintensity within 10-fold of the strongest array signal in a particulargroup. In addition, sense oligos printed elsewhere on the array (i.e.,outside the group of 200-300 oligos being tested) that exhibitsignificant cross-hybridization are also resynthesized. This first testconfirms that the oligo can yield a strong hybridization signal. Thesecond test is similar to the first, except that antisense oligos areprepared synthetically. This analysis verifies the actual sequence ofthe oligo. The synthetic test is especially useful for identifying bothformatting errors and poor oligo synthesis.

[0111] I. Introduction-Use of Antisense Oligo Mixes Because of therigorous testing that has already been performed on Clontech oligocollection, hybridization with an Antisense Oligo Mix yields highlypredictable results. In practice, adequate signal (an intensity within10-fold of the median array signal) is typically observed for at least98% of all printed oligos on our premade or custom arrays. As a result,Antisense Mixes provide a highly effective quality or calibrationcontrol. Antisense Oligo Mixes are available for human, mouse, and ratarrays and can be used with any BD Atlas Glass Microarray of thecorresponding species, regardless of array content. Thus, they arecompatible with our premade arrays, custom arrays, or arraysmanufactured using BD Atlas Ready-to-Print Long Oligos. Each oligo in anAntisense Mix is tagged on the 5′ end with a primary aliphatic aminogroup, and can be labeled directly with Cy3, Cy5, or othermonofunctional N-hydroxysuccinimide-ester-activated dye.

[0112] II. List of Components

[0113] Store Oligo Mix and Labeling Buffer at −20° C.

[0114] Store DMSO tightly capped at room temperature, in the dark.

[0115] Each BD Atlas Antisense Oligo Mix is sufficient for 5 labelingreactions.

[0116] Notes:

[0117] DMSO may be safely stored at −20° C.

[0118] Depending on the complexity of the oligo mix, one labelingreaction may yield sufficient probe for

[0119] multiple hybridizations.

[0120] 50 μl BD Atlas™ Antisense Oligo Mix (50 μM)

[0121] 100 μl 2× Fluorescent Labeling Buffer

[0122] 500 μl DMSO

[0123] III. Additional Materials Required

[0124] Fluorescent dye The fluorescent labeling protocol is optimizedfor use with the following dyes:

[0125] Cy3 Mono-Reactive Dye Pack (Amersham Pharmacia Biotech #PA23001)

[0126] Cy5 Mono-Reactive Dye Pack (Amersham Pharmacia Biotech #PA25001)

[0127]  Each pack of Amersham Pharmacia dye contains five vials. Eachvial contains sufficient dye for four Atlas Glass labeling reactions.Please disregard instructions for use and protocol information suppliedwith these dyes. Complete procedures are supplied in this User Manual.Other mono-functional, N-hydroxysuccinimide-activated fluorescent dyesare also compatible.

[0128] MERmaid Spin Kit (Bio 101 #1105-600)

[0129] For Purification of Labeled Oligos

[0130] 0.5 M EDTA (pH 8.0)

[0131] 3 M Sodium Acetate (pH 5.3)

[0132] 100% Ethanol (Avoid denatured alcohol; we recommend Spectrum#E1028)

[0133] 70% Ethanol

[0134] Quartz cuvettes

[0135] UV/Vis spectrophotometer (Optional: a scanning spectrophotometerwill assist you in troubleshooting probe quality.)

[0136] IV. Antisense Oligo Labeling

[0137] A. General Considerations

[0138] The optimal amount of labeled Antisense Oligo Mix to use in yourhybridization depends on the complexity of the Oligo Mix and is notaffected by the oligo content on the target microarray. The approximatecomplexity of each Oligo Mix is indicated by the product name. Forexample, the Human 8K Antisense Oligo Mix contains approximately 8,000oligos. For each hybridization, you should plan to use approximately 25pmol of labeled Oligo Mix for every 1,000 oligos in the mix. Thus, forthe Human 8K Mix (provided at 50 μM), approximately 4 μl of the initialOligo Mix are required per hybridization:

[0139] 25 pmol×8=200 pmol

[0140] 200 pmol=4 μl

[0141] 50 μM

[0142] The procedure below is designed to label 10 μl of Oligo Mix, andis suitable for 2 hybridizations using the Human 8K Mix. For mixes ofhigher complexity, you may scale the procedure up, as necessary.

[0143] B. Antisense Oligo Re-suspension

[0144] Follow the steps below to prepare the Antisense Oligo Mix forlabeling.

[0145] 1. Transfer 10 μl Antisense Oligo Mix to a 0.5-ml tube.

[0146] 2. Precipitate the Antisense Oligo Mix by adding 1 μl 3M SodiumAcetate (pH 5.3) and 27.5 μl of ice-cold 100% ethanol. Vortex gently.

[0147] 3. Place tube in a −20° C. freezer for at least 1 hr.

[0148] 4. Centrifuge tube at 4° C. for 20 min at 14,000 rpm.

[0149] 5. Carefully pipette off supernatant, and air dry pellet briefly.

[0150] 6. Wash pellet once with 70 μl of 70% ethanol. Flick tube gentlyto wash.

[0151] 7. Centrifuge tube at 4° C. for 5 min at 14,000 rpm.

[0152] 8. Carefully pipette off supernatant, and air dry pellet.

[0153] 9. Add 10 μl 2× Fluorescent Labeling Buffer. Do not substitute adifferent buffer. Proceed directly with Section C.

[0154] C. Fluorescent Dye Coupling

[0155] Proceed with the steps below to couple fluorescent dye to theantisense oligos. IMPORTANT: Use only the DMSO provided or Atlas GlassApproved DMSO.

[0156] 1. Prepare a 5 mM stock solution of fluorescent dye by adding theappropriate quantity of DMSO directly to the dye container. If you areusing Amersham Pharmacia Cy3 or Cy5 reactive dye, open one pouch of dye.Each pouch contains one tube of dye residue sufficient for four labelingreactions. Add 45 μl DMSO directly to the dye vial. Vortex and brieflyspin down.

[0157] 2. Add 10 μl of the DMSO/dye mixture to your 10 μof oligo mix.Mix well and place the tube at room temperature in the dark or wrappedin aluminum foil. Incubate at room temperature for 30-60 min.

[0158] Note: The remaining DMSO/dye solution can be stored tightlycapped at −20° C. for at least 1-2 months without noticeabledegradation.

[0159] 3. Add 2 μl 3M Sodium Acetate and 50 μl 100% ethanol; vortex.

[0160] 4. Place tube in a −20° C. freezer for 2 hr to precipitate thelabeled probe.

[0161] 5. Spin tube at maximum speed in a microcentrifuge for 20 min.

[0162] 6. Carefully pipette off supernatant, and wash pellet once in 70%ethanol.

[0163] 7. Dissolve pellet in 100 μl Deionized H₂O.

[0164] D. Probe Purification

[0165] Follow the steps below to purify your labeled antisense probeusing the MERmaid Spin Kit. Each SPIN Filter is suitable for purifying˜1 nmol of labeled antisense oligos. For larger-scale purifications,perform the following steps in parallel, for each 1-nmol equivalent.

[0166] 1. Completely resuspend GLASSFOG Bind solution, and transfer 400μl to a SPIN Filter.

[0167] 2. Add the entire 100 μl of probe and mix with rotation at roomtemperature for 5 min.

[0168] 3. Centrifuge sample at 14,000 rpm for 30 sec.

[0169] 4. Add 500 μl MERmaid Spin Ethanol Wash, and centrifuge at 14,000rpm for 30 sec.

[0170] 5. Repeat Step 4 two times.

[0171] 6. Empty catch tube and centrifuge at 14,000 rpm for 1 min to drypellet.

[0172] 7. Transfer SPIN Filter to an elution catch tube.

[0173] 8. Add 50 μl MERmaid Elution Solution, and resuspend GLASSFOG byflicking the tube. Centrifuge at 14,000 rpm for 30 sec.

[0174] 9. Repeat elution Step 8 using 50 μl MERmaid Elution Solution.

[0175] 10. Add 150 μl of H2O to the catch tube to bring the total volumeto 250 μl.

[0176] E. Analysis of Probe Quality

[0177] To assess the quality of your labeled probe, analyze your entireprobe using UV/Vis spectrophotometry. Ensure that your cuvettes andspectrophotometer can accommodate the small volume of labeled probe(−250 μl). The measurements described in this section assume the use ofcuvettes having a 10-mm path length (such as Sigma #C1918 or #9917). Forthe most rigorous analysis, we recommend using a scanning instrument toread the full absorbance spectrum from 200-800 nm for both Cy3 and Cy5probes, using elution buffer as a blank. The instrument absorbancereadings for Cy3 (A550) or Cy5 (A650) probes are typically around 0.06,when starting with 500 pmol of Antisense Oligo Mix. Use the formulabelow to determine the optimal amount of probe to use in thehybridization (Vopt) based on the absorbance reading and complexity ofthe probe mixture. This optimal amount of probe assumes use of the BDAtlas Glass Hybridization Chamber, using a hybridization volume ofapproximately 2 ml. If you are using a different final volume, theamount of probe will need to be adjusted proportionally, so that thefinal oligo concentration is not altered. Further optimization may berequired for other hybridization techniques.${{Vopt}\quad ({µl})} = \frac{M \times 1.25}{A\quad \lambda}$

[0178] Where: M=Probe Complexity Multiplier

[0179] (e.g., M=8 for the Human 8K Mix; M=5 for the Mouse

[0180] 5K Mix)

[0181] Aλ=instrument absorbance reading at 550 nm (Cy3) or 650 nm (Cy5).

[0182] Proceed with microarray hybridization.

Example 3

[0183] In this example, a schematic is provided in FIG. 5 showing Witheach new lot of microarrays printed, several microarrays (some from thebeginning, middle, and end of the printing) are hybridized using anantisense oligo calibration mixture. Following quantitation, theresulting lot-specific calibration values are averaged, such as thoselisted on the Clontech web site. These values can be calculateddirectly, or can be imported into appropriate software, such as BDAtlasImage™ Software, which will automatically calculate standardizedarray signals, yielding the most accurate and meaningful arraycomparisons. This standardization protocol is ideal for databasegeneration, as it allows statistically significant data to be generatedfrom microarrays printed at different times. FIG. 5 describes theimportance of array calibration to generate accurate, meaningfulresults.

[0184] Panel A first illustrates the calculation of lot-specificCalibration Standards for a target gene. Then, two different RNA samplesare analyzed for target gene expression differences using two arrays—onefrom each lot (Panel B). Without calibration, the target gene appearsup-regulated (Raw Signal, Panel B). Our practice of gene standardizationdemonstrates how the lot-specific value corrects for typical printingvariations across lots (Calibrated Signal, Panel B). In this case, arraycalibration shows an insignificant difference in gene expression.

[0185]FIG. 5 demonstrates a generalized approach to provide moreaccurate expression data, such as with the use of calibrated BD Atlas™Plastic Microarrays.

[0186] Panel A. After printing each lot of BD Atlas Plastic Microarrays,sample arrays from the beginning, middle, and end of the printing runare hybridized with a mix of synthetic 33P labeled antisenseoligonucleotides corresponding to all genes on the array. Then, theintensity of each hybridization signal is quantitated by phosphorimagingand averaged. Average antisense intensities are calculated for eachgene, as shown above for hypothetical Gene Calibration Standards arethen calculated for each array lot relative to the initial printing run.All genes in the first printed lot (Lot 1, as shown) are assigned aCalibration Standard of “1.0”.

[0187] Panel B. After normalizing arrays based on the overall signalintensities from all genes on the array, experimental intensities forGene X can then be compared using calculations that correct for arrayprinting variations between lots. Without this correction, geneexpression comparisons are less accurate and less reliable.

[0188] The above results and discussion demonstrate that novel andimproved methods of producing aRNA from an initial mRNA are provided.The subject methods provide for an improvement over prior methods ofproducing aRNA in that the cDNA synthesis step is not required. As such,the subject methods represent a significant contribution to the art.

[0189] All publications and patent applications cited in thisspecification are herein incorporated by reference as if each individualpublication or patent application were specifically and individuallyindicated to be incorporated by reference. The citation of anypublication is for its disclosure prior to the filing date and shouldnot be construed as an admission that the present invention is notentitled to antedate such publication by virtue of prior invention.

[0190] Although the foregoing invention has been described in somedetail by way of illustration and example for purposes of clarity ofunderstanding, it is readily apparent to those of ordinary skill in theart in light of the teachings of this invention that certain changes andmodifications may be made thereto without departing from the spirit orscope of the appended claims.

1 1090 1 80 DNA Rat 1 ctctcacctt ggtcatgatc ttgagctgca tgatctgctgccggatggtc atatccggct 60 tggtgatgtc caactctccc 80 2 80 DNA Rat 2ctctcaccca acttctttgc tgtcttcctg ggctttgttc agaggatcag ctggctgaac 60acagtttgcg tcactctccc 80 3 80 DNA Rat 3 ctctcaccgc ccagcacttg agtgatcaatatggcaatgg tgaaagaggt cattccaaaa 60 gctgtcataa acactctccc 80 4 80 DNARat 4 ctctcaccgc tctacataga cctgactgga gcttgggtgc gatgtggagg tactggtggg60 gatgtagtct gtactctccc 80 5 80 DNA Rat 5 ctctcaccta cagcactggcgtctgccgcc cgcgcttcga ggagtcctta gaagacttgt 60 gaaatttccc ctactctccc 806 80 DNA Rat 6 ctctcacctc agggtctcta ttggcctgaa ggaggaccac caacatcctctggtagtacc 60 ctgaggtatc ccactctccc 80 7 80 DNA Rat 7 ctctcaccgccatatggatg gctttgagtg tttcccgctg gggctcatcc ttgttcctta 60 acttttggtaggactctccc 80 8 80 DNA Rat 8 ctctcacctt gatgggacgc tccagctcag gcttcttaaagcggagccac atcatgccga 60 tgatggccag ggactctccc 80 9 80 DNA Rat 9ctctcaccat gaggagccgg ctgcccgaga ggagaccacg gtccttccct cccccacccc 60cctctcccct ctactctccc 80 10 80 DNA Rat 10 ctctcaccgt ggtttcaaatatcgaagcca gaatgcatgc ctgtctcagg gtcaccacgc 60 cagagcccac cgactctccc 8011 80 DNA Rat 11 ctctcacctc agtccctggc atggattata acattatttt caaaataataaaaaattaaa 60 aattatacat atactctccc 80 12 80 DNA Rat 12 ctctcaccttccccgatgat aacagtggct gtgttgggct ggcccaggcg ggctccaaac 60 ttggggttgctgactctccc 80 13 80 DNA Rat 13 ctctcacctc agattcttca gtgcttctatttttctagct ggcttgacca gtatgttgac 60 ctgcagtcgt ttactctccc 80 14 80 DNARat 14 ctctcaccct catctggtgg tggtaagatt ccaggggtgg gcattcttat ggctttttga60 aggattccct ggactctccc 80 15 80 DNA Rat 15 ctctcaccca ggcccctggtctattcactc tcttttgaag agagagcctc cacctcctgt 60 agcagcacat ggactctccc 8016 80 DNA Rat 16 ctctcaccat gagagcagtc caagctgcct tgttctgggg caaagatttctgggcacttg 60 atggcttcac acactctccc 80 17 80 DNA Rat 17 ctctcaccttccttgatctt tgagaaactt ctctttgtct tttgacatat tggactagga 60 ctagaccaggagactctccc 80 18 80 DNA Rat 18 ctctcaccga gtggagagga agtgctctagctcagaggca ggagagaagt ccttgcccac 60 tcccaagcat gaactctccc 80 19 80 DNARat 19 ctctcaccaa aatctcgtca gggaacacag ccctgtcttt cgttactgtc tttgaacatg60 ataccacttg atactctccc 80 20 80 DNA Rat 20 ctctcaccgg gaggcagccatgacaaaggc agtggtgttg caggactgga tggtaccgga 60 actgtcacag ccactctccc 8021 80 DNA Rat 21 ctctcacccc ttcacagaga ccaccaggtt gtagtacgga ttctttattggatccaatac 60 ctgagaacct tcactctccc 80 22 80 DNA Rat 22 ctctcaccttcttgcaaagt aaatgatcat cccaatggcg ggtattacca aggaggatgc 60 aaagtagagtgcactctccc 80 23 80 DNA Rat 23 ctctcacctg attgtcaagc aacaaaggggtgactagagg aaagacttca ctactgtagg 60 aaggttgaag agactctccc 80 24 80 DNARat 24 ctctcaccgc agcacactgc atggtagcgt tttctccaac agtgatatca gcattgattg60 gggccaaaat gaactctccc 80 25 80 DNA Rat 25 ctctcacctc tatcaaaaacgtttgcagtt tgtaatttgc aagttccttt ggtttaccaa 60 gcaaatatta agactctccc 8026 80 DNA Rat 26 ctctcaccca ctttgctcct ccccagaagc gcccccacat cttcagcctgtatttccatt 60 tcatacaatg agactctccc 80 27 80 DNA Rat 27 ctctcaccacccaagaactg ggctggggta tgagcgaggg gagctctccc tttagagagt 60 ccaggggtgacaactctccc 80 28 80 DNA Rat 28 ctctcaccct tcctcaggac agagggatgtgcttggtctt tgttgctggc cttaagttct 60 tttacagtct ctactctccc 80 29 80 DNARat 29 ctctcaccca caaataagct catacagtgt tcttcttgga ggtagtaata attgaaaagc60 accaacagaa aaactctccc 80 30 80 DNA Rat 30 ctctcaccaa aagaccaaccagcagcagca caaagagatg aatgaacctg agacgtggaa 60 acccgaggag tgactctccc 8031 80 DNA Rat 31 ctctcaccga ctttctagag agaaataaat cacttaagca gaaggtaggtgtaaatcaca 60 gccctaaagc ctactctccc 80 32 80 DNA Rat 32 ctctcaccccacacacatta agaacttcac cggcgggtag tcattgatac agctgtgctg 60 gaaccccaaggcactctccc 80 33 80 DNA Rat 33 ctctcaccct aagaataagg ctagagaatgtcgccctcgt ccacgtgagc agagctggca 60 tccagtcttg atactctccc 80 34 80 DNARat 34 ctctcaccat gtccctcacc tggaatgtgg cagccagaaa tagccattgt ctctcccttt60 gctctggagt caactctccc 80 35 80 DNA Rat 35 ctctcacctt gcctcttccggtacctttta tatatgaaca caaatagaat tgccaaaccg 60 gggaggatca gcactctccc 8036 80 DNA Rat 36 ctctcaccgc aaaagagttc ccagttatgc tgtcatccag cagcggcagggtctccaggc 60 aggcggttcc aaactctccc 80 37 80 DNA Rat 37 ctctcacccaagtaggttcc tttgtggcac ttggtgcagc agatggaatt attctttgga 60 tgggcatactttactctccc 80 38 80 DNA Rat 38 ctctcaccgg agtacacctg cagacacttctcctttagca cacaagtggc agtgaagtta 60 gagccacgct ggactctccc 80 39 80 DNARat 39 ctctcacctt gtcatagtct tcaacagaga cctgtgtggg gttccccggg atccctgccg60 gagtggtcct ggactctccc 80 40 80 DNA Rat 40 ctctcacccc cgagggatcgaaaggagaca ggcacccgtc gagtgcacgt ttgacccctc 60 aggtgtttct tgactctccc 8041 80 DNA Rat 41 ctctcaccga gcagggaggc aatgaggata agatgagcag ggcctggcccgggggcgccc 60 cctgtagggc ggactctccc 80 42 80 DNA Rat 42 ctctcacccacatttcgagc aaatcacgtc aattttggat gcttgtcttc atctcggaca 60 taaaactccagcactctccc 80 43 80 DNA Rat 43 ctctcaccct ctactctaac ttccccagtcaagtcaataa atatattaag cttgtgtccg 60 ggtcaagcca caactctccc 80 44 80 DNARat 44 ctctcaccca gagcggctat tcccaggcct cccaggatcc ctaaggccag ggctagcgtt60 cccagcccag acactctccc 80 45 80 DNA Rat 45 ctctcaccta taccgttacattataggtgc cactgtcatc tcttgtcaga ttctttatct 60 tcaagtctcc atactctccc 8046 80 DNA Rat 46 ctctcaccat cagagactca gagtaggcct gcatggcctc cttatcctgcgagccctttt 60 tactaccact ccactctccc 80 47 80 DNA Rat 47 ctctcaccgctctaacaatc actaaagatc cattcacaga ataatttaat tcttcaaagt 60 cctttttagtgcactctccc 80 48 80 DNA Rat 48 ctctcaccaa gaacaacacg gccctagataggcacacagg caagaaatcc aataaagagg 60 cagaagaaac agactctccc 80 49 80 DNARat 49 ctctcacctt tgctagtccc aagggtctag accctcgtcc tgatgctccg accgtgagaa60 aactgtggga gtactctccc 80 50 80 DNA Rat 50 ctctcaccgg agtgtgtgtggggattctga gatccaagtc cttcacacat taagtctaca 60 tgatggatgc tcactctccc 8051 80 DNA Rat 51 ctctcaccca gtgagtggtt tgccactcct ggatgttctc tgtatcgaggtgaactgatg 60 tacagcaaag caactctccc 80 52 80 DNA Rat 52 ctctcaccggtgagcatcgt ggcctggtgg aggtgtcaca tgaccagagt tcagtttacc 60 caggattttgtgactctccc 80 53 80 DNA Rat 53 ctctcaccct gcaggctgct gtggtcatcgatgtcgctga tgctggtggt actgatgttg 60 tccacttctc caactctccc 80 54 80 DNARat 54 ctctcacctg cagcttctcc gtcagctcac gtcgacggtt ccgacacttg gcagcggcca60 gcttgtttct ctactctccc 80 55 80 DNA Rat 55 ctctcaccgt agtcgtgaagagaaagactg ccagggctcc gaccgtatcc tgccgctgcg 60 taagagtcgt cgactctccc 8056 80 DNA Rat 56 ctctcaccga aagcgcggcc cgggggcgcg aagccaccac cgccgggggccccagcagca 60 ccagcgacgc tcactctccc 80 57 80 DNA Rat 57 ctctcaccagcccgtttgtc agccgcagat tgaaacctcg gttgctcttc gtcgctctcc 60 acctcgatgtcaactctccc 80 58 80 DNA Rat 58 ctctcaccaa gctcttgcta tacttgggatggcaatactt cccaagttgc catctagtgt 60 ctttccaggt caactctccc 80 59 80 DNARat 59 ctctcaccgt ccttttatca gattctggaa aatcacctca aaaaaaaaaa aaaaatccca60 caaacatgaa ccactctccc 80 60 80 DNA Rat 60 ctctcacccc caaagacagcagaagtagga ggttaaagaa aaaatctgga cacagtgagt 60 tcagtgatct gaactctccc 8061 80 DNA Rat 61 ctctcaccca tttcagagcc aagaaaggaa gccaaattcg gaaggcctcgaatgacatca 60 agattgaccc tgactctccc 80 62 80 DNA Rat 62 ctctcaccgtagaatggtag tacccggctg agtggcagct gcattggtca tggttaatgt 60 ctgcaggccctgactctccc 80 63 80 DNA Rat 63 ctctcaccag tttagcttta tgttgtatcacagccaccat gatgctggac ctagctcact 60 caggagggca agactctccc 80 64 80 DNARat 64 ctctcaccat tcagggtggg gatgatggtc cctcagcttc acaaacacac caacaatgcc60 ggttccacat ttactctccc 80 65 80 DNA Rat 65 ctctcaccct gcttctgaagagaaatgcag tccagctaca aaggaggctg cggggccaaa 60 gttacatcta tgactctccc 8066 80 DNA Rat 66 ctctcaccca agtatattta gatatgttca ctttctccaa tttcactgtggaagcatctt 60 caataagagg caactctccc 80 67 80 DNA Rat 67 ctctcacctgctgcaggctg ggtggaaagc catagtccag gaataccatg tccttaaggg 60 aggccactgtcaactctccc 80 68 80 DNA Rat 68 ctctcaccga ggtgatggtt gggtgggggtccggagggct tctagggtgt gcttggaatc 60 tcgaccctcc agactctccc 80 69 80 DNARat 69 ctctcaccat gactgtaatt ggaccggttt ctctctcctc aggttccggc tgttattttg60 gctctgtcgc tgactctccc 80 70 80 DNA Rat 70 ctctcaccgg caccagctccttgaggcgtg agtagcagcc gttcatgtcg tagagcagaa 60 cgttcacctg ctactctccc 8071 80 DNA Rat 71 ctctcaccgg ggtgcgagtc cagggcgatc tgcaggtcca agatataatcgatgacgtgc 60 tgcaggattt ccactctccc 80 72 80 DNA Rat 72 ctctcaccggggtgcgagtc cagggcgatc tgcaggtcca agatataatc gatgacgtgc 60 tgcaggatttccactctccc 80 73 80 DNA Rat 73 ctctcaccag cactccacca ccttgccaaaggtgccttca cccaggttcc ccacgatctc 60 atatcgctct tgactctccc 80 74 80 DNARat 74 ctctcaccaa gggctccctc tcccagttac agtgaagtca aaatcattag ggtccaagga60 ggtgtctccc acactctccc 80 75 80 DNA Rat 75 ctctcaccga gaaagcctgacacagatatt cttcggcagg tgcacatcca gatgtttcca 60 ttgggcttgg agactctccc 8076 80 DNA Rat 76 ctctcaccct ctgccagaag ttccctgcca tggaatactg cttggcctgatacaaaagca 60 ccagccggca aaactctccc 80 77 80 DNA Rat 77 ctctcaccctcaggtttgga tggctctcca atgggggcaa ttgggttggg aaagtcaagg 60 agacccacatctactctccc 80 78 80 DNA Rat 78 ctctcaccac aggtcaacat cccgcacgtctgtaggggtg gtggcttggt ccggatcttc 60 cacagacttg gaactctccc 80 79 80 DNARat 79 ctctcaccgc aacagtttac gggcgcttgc tcttctagcc aaggcctagg ctggttcccg60 ctaagagcag ctactctccc 80 80 80 DNA Rat 80 ctctcaccaa ttgggactttccgcaactgt gaggccattt gccaacagct aacgtggtga 60 ggggctgagg ttactctccc 8081 80 DNA Rat 81 ctctcaccgg gcactggttt agtcacatcc tggaagaaag ggtgagccagggctgctttg 60 gctgaaatcc gcactctccc 80 82 80 DNA Rat 82 ctctcaccgctgtaattcta gtacatgggt tgaataagaa caagccttgt atgagctcaa 60 gcaggtcgtctcactctccc 80 83 80 DNA Rat 83 ctctcaccca ccccaagaag agaaactgggcttgctaaca gcagaggagt ctcaccacca 60 agcaggtcac caactctccc 80 84 80 DNARat 84 ctctcaccaa agtcgaaatt ccacttgcgc tggctcgctt cttccatatc tcggcagtgc60 ttctccaagt ccactctccc 80 85 80 DNA Rat 85 ctctcaccat atcacggtggccctgctctt cagctaagtc taccggcagg cggccccagg 60 cgtcacacac atactctccc 8086 80 DNA Rat 86 ctctcaccca tggcggaaag cactcggcca gactgtggtc gaaacaggcacagaattggc 60 ttcccaagag ctactctccc 80 87 80 DNA Rat 87 ctctcacccaggtttgctag gggtggtctg aaccttagaa ttagatttgc caggagttcg 60 gcccggggtccgactctccc 80 88 80 DNA Rat 88 ctctcaccat acgtgcaaat tcaccagatggcatctttac tacgcagctg tactcctgtt 60 ctgggattcc aaactctccc 80 89 80 DNARat 89 ctctcaccag tcctcgccaa tgctggtgta gatacgagga agctggctgg ccaccggccg60 gaagaggata cgactctccc 80 90 80 DNA Rat 90 ctctcaccat cttcatccgcagccagcagg cacaggacca cgttgtccgg gtctacgttg 60 agcagcttgg ctactctccc 8091 80 DNA Rat 91 ctctcaccaa gttttttaaa caaatcactt atgctgtgat aaaccaaagatgagtaactg 60 catccaacta atactctccc 80 92 80 DNA Rat 92 ctctcacctccttcaagtcc ttggtggtga tctcggagct ggtgtccact gccgcgtctg 60 acatggtggggcactctccc 80 93 80 DNA Rat 93 ctctcacctt ttgctccata ggaaaagaccgcaccgaaga tatcctcatg ataacgtttc 60 tggctcttga gcactctccc 80 94 80 DNARat 94 ctctcacctg attaggatcc aaaatgaact caaaggtctc attccacaca gggtttatat60 cattattgaa gtactctccc 80 95 80 DNA Rat 95 ctctcaccct gagggtgtatgaaaccccgt tgatgaggaa ggtgacgttt ggcatcatgt 60 tgagagtggc acactctccc 8096 80 DNA Rat 96 ctctcaccac cctgagaggt gccagcctcc tgtgctttga gagggtcgtgtcttactgct 60 gaagtagaat gaactctccc 80 97 80 DNA Rat 97 ctctcacccttgttcctggc agggttctaa aatgaggata tggtgaatag gaatggtact 60 cagagtgaagctactctccc 80 98 80 DNA Rat 98 ctctcaccaa tggctttggc cagtaaggttttcccacagc caggcggtcc atagaaaaga 60 acacctttgg aaactctccc 80 99 80 DNARat 99 ctctcaccta gccaattcct cctctccctg accccaagac acgtgagcaa ctgctaatga60 aaagcagtaa acactctccc 80 100 80 DNA Rat 100 ctctcaccgg gaagaaaacggatgaaggaa gatggaaaat cataacagca agtctgatgc 60 aggactgtga agactctccc 80101 80 DNA Rat 101 ctctcaccta cattttgagc gtttcttctc attctgtcccaacgccacat aagccaggtt 60 tgtgggcgca tgactctccc 80 102 80 DNA Rat 102ctctcacccg atgaggaaag gcagatccgg cttggacagc ctcagcaacc gtatcatgag 60tgctctgtta ttactctccc 80 103 80 DNA Rat 103 ctctcacccc aatgatggaccctccacaca tatggttctg agaaaccaac ttcacttgca 60 ggctgacctg ccactctccc 80104 80 DNA Rat 104 ctctcacctt catctgccaa ggacagggtt gatcactgtgaatggacatg aaaccagtca 60 accagtctca gaactctccc 80 105 80 DNA Rat 105ctctcacctc agctgccgag cctgagcggc agcagaagat gtgatcgtgg ataggggtca 60gcttgtcagt caactctccc 80 106 80 DNA Rat 106 ctctcaccgg aaccgccgatcgcaaagggc tgtcgaatta gcatccctcc catggttcca 60 tacacctggc ccactctccc 80107 80 DNA Rat 107 ctctcacctc cactttcacc caaccgtctt tcttcatgtggtacatgttg acaactcctc 60 cagaatagct gtactctccc 80 108 80 DNA Rat 108ctctcacctc ggtgcccaga ggctccagct tctggcggta ggcctcgacc tcctcgttcc 60acttctcctg gaactctccc 80 109 80 DNA Rat 109 ctctcacccc tgcacaccctccgcgagagg ggccagtctg ctgcgcaggt cttccaggtt 60 ctggtcgatc gtactctccc 80110 80 DNA Rat 110 ctctcaccgg aggctcctca gacagttgcc ggagagattcataaccgcca cattgaaaag 60 gccagagaag gcactctccc 80 111 80 DNA Rat 111ctctcaccac aggaagtccg ggcagtttgc ttggtctttt gttgttattt tggcgatgtc 60gatgtcatta gaactctccc 80 112 80 DNA Rat 112 ctctcaccgc ctggtggcccagccagatat ctcggcgtat acagccatcc agggcaggaa 60 cgagcggaaa ccactctccc 80113 80 DNA Rat 113 ctctcaccaa acagcaatat cttctcattc actgtctttaacatgaatgc cgtgctccct 60 tcatactttg caactctccc 80 114 80 DNA Rat 114ctctcaccag atcgtatgag ggcccaagat agaatcttac aggcggtggg gtggggtgaa 60atttcactgt agactctccc 80 115 80 DNA Rat 115 ctctcaccta aaaacagtctacgtacatta tttacagtca ggtcagggag ggaagccatc 60 ctgctcagct ttactctccc 80116 80 DNA Rat 116 ctctcaccta ctagagagaa cactgggtta caggactacatgcagctgag agccagcgca 60 caagagcctt tcactctccc 80 117 80 DNA Rat 117ctctcaccgc tgggaggtca tctggtataa attgtcactg ctgtagggcg tcctcaggag 60cagggcctgg ctactctccc 80 118 80 DNA Rat 118 ctctcaccgc taatgttcttgaccggctcc aaggatggct tgggctcagg gtcattgaga 60 agaggcaggg tgactctccc 80119 80 DNA Rat 119 ctctcaccct gaggacctgg ggctggagca tttgcggttgttgctgatct gtttcaggac 60 cctgccactg tcactctccc 80 120 80 DNA Rat 120ctctcaccgc aggacgaccg ctgccacgag caggagcagc caccggggct acagccggga 60cagacacact agactctccc 80 121 80 DNA Rat 121 ctctcaccag gcgccactctcggactggag gaacgaggcg ttgagggcat cgtcgtagaa 60 ggtcgtttcc atactctccc 80122 80 DNA Rat 122 ctctcacctg ctgagcccct ggtggtcacc aaggcttatcgctgcgccag cactggcagg 60 gatggtggct acactctccc 80 123 80 DNA Rat 123ctctcacctt ctggaaaata aaacatagaa taacaattaa aaattaagag tccaaccaac 60acgggttgtt ctactctccc 80 124 80 DNA Rat 124 ctctcaccca tccttggactgctttatgag atcaaacaaa ggtgaatcag aaagccatgc 60 aagggattcc atactctccc 80125 80 DNA Rat 125 ctctcacccc ttctatctag gcttagagta ggctggataacatgaagaaa gtgacatggt 60 gtgggaaagt ggactctccc 80 126 80 DNA Rat 126ctctcaccat caccaggatc ggacatggtg cctgtggctc tgagtgtctc gatatcgctg 60tggatcggcg ccactctccc 80 127 80 DNA Rat 127 ctctcaccga ccatttcccccaggtcatgc gtactaagtg tctttatgta catttataca 60 tttttaagtg ctactctccc 80128 80 DNA Rat 128 ctctcaccaa gcgtagcacg cgcagaaaca ggactcagaaagaacggtgc taacagagct 60 ccaggtgcag gaactctccc 80 129 80 DNA Rat 129ctctcaccgc tgtcactcat agatccagtt ctgcgcacag ctcttgtagt ccaccagctc 60ctcaggctga aaactctccc 80 130 80 DNA Rat 130 ctctcaccgg ggtatttcgaacagattgct gaggtggctg agagtcaaag ggcatcatca 60 tttttggtct caactctccc 80131 80 DNA Rat 131 ctctcacctg tcttctttgc caaacaaggg aaggccaaccaggaaaagta cggcaggatc 60 tctaacgaac acactctccc 80 132 80 DNA Rat 132ctctcaccgc cagttgatgc tctgcacatc atcgcgcagc cggcagcgaa gctgtagcag 60gtcaccaggg tgactctccc 80 133 80 DNA Rat 133 ctctcaccga gcatgtccaggtgggtctca ggactggcag ggagccgcaa cttcatgtct 60 gtgccggtac acactctccc 80134 80 DNA Rat 134 ctctcacccc cagcatcccc cagaggaaca cagtggatcacacactttgc cctctgccaa 60 gcattctccc agactctccc 80 135 80 DNA Rat 135ctctcaccag acagggatgg ggtcctcagg gttgaactca aaagggttgt ccataaaggc 60agccatgatg gaactctccc 80 136 80 DNA Rat 136 ctctcaccgc aggctctgctgcagccagag accctaggct cctcctccct cgagatgagc 60 tttccgactc gaactctccc 80137 80 DNA Rat 137 ctctcaccaa acattcacag agaaaggtgg aatggagaagaatgaggaac aggcagaact 60 tgctgagaat ttactctccc 80 138 80 DNA Rat 138ctctcaccat tcacagagag atttgaggga acctcctgct gcctggagcc tccagacaaa 60tcctggatgc tgactctccc 80 139 80 DNA Rat 139 ctctcaccgc cattggatggatcttgaaac cgtagagcct gggcacaaac tggttggccg 60 gccgcttggg ccactctccc 80140 80 DNA Rat 140 ctctcaccgc agacaagctc tctcttccct ggagggtctgacagaatggc tgctatttgc 60 tgggtgatgg tgactctccc 80 141 80 DNA Rat 141ctctcaccct gaacaattgt aggggagatg caactggagc catcaaacac agcatctttg 60agtccgaaac caactctccc 80 142 80 DNA Rat 142 ctctcacctg ttgccaacattaaggaaagc agggctgatg ggctgaggcg agatggacga 60 agaccggcct tgactctccc 80143 80 DNA Rat 143 ctctcaccgg ggtctgtgcc atagcggtac ccagagctctggttcaggct gccgtccctc 60 tcctccgtca gtactctccc 80 144 80 DNA Rat 144ctctcacctc atctgagctg tgaactcctc atcaaaatac ctggtgtcgg tctcagaggt 60gacctggggc ttactctccc 80 145 80 DNA Rat 145 ctctcacctg ggacagtcacaagagacgct agagagaaat gaccaggaac acccagggaa 60 gtttggaagc atactctccc 80146 80 DNA Rat 146 ctctcaccca ctctcatcag gcgggttcag tttccgcagtttatgctgcc gaatctcacg 60 tactagtgtg taactctccc 80 147 80 DNA Rat 147ctctcaccat ttgatctttc gcagactgca ctattggcgt gaagaacagt gtggggtgct 60ccacggaaag tcactctccc 80 148 80 DNA Rat 148 ctctcaccga gcctgtttcgtgtctactgt tctagaaggc aagtcacact tattccctac 60 taggaccata ggactctccc 80149 80 DNA Rat 149 ctctcacctg ccttgcccca acccacctct ctccatctatggcaatgcgg caggggacgg 60 ataggcctga ctactctccc 80 150 80 DNA Rat 150ctctcacctg agctacacaa atctcatgca atgctgctaa gtcacgagac aggtcagttc 60tagaatgttc tgactctccc 80 151 80 DNA Rat 151 ctctcacctt cgcttattgagatggacacc tggcttgcat tctggaccac agcgccatcc 60 ttcatccagt gtactctccc 80152 80 DNA Rat 152 ctctcacctg aggctggtgt tggtggcatt caatatacttgcttctacca tattggaagc 60 tacagacaga taactctccc 80 153 80 DNA Rat 153ctctcaccgc caaatagaaa agtcaccgat aacgtcacga ttgtgtagta atggccagaa 60gagagttcat agactctccc 80 154 80 DNA Rat 154 ctctcacctt cctcagaccgcatcagagct tcattggcca tcgttgcttc ctctttcatg 60 tgcaggaccc tcactctccc 80155 80 DNA Rat 155 ctctcaccaa ctgtgcactg gagtccaatt taatctgtttgccagggtcc aatccaagac 60 cccgcgctgc acactctccc 80 156 80 DNA Rat 156ctctcaccct gcaagaatat ctccttcctc tgctttgcta ccatccaggt ctggtatagc 60tatagaggag acactctccc 80 157 80 DNA Rat 157 ctctcacctc aattccaggcaagtacgtgt caatcaaggc atcaagaaat ttaacaatga 60 gctcagcata gtactctccc 80158 80 DNA Rat 158 ctctcaccag aggcggctgt gggcagggta ccagtcccggaacggctccc agctggggct 60 ccgcagtagc gaactctccc 80 159 80 DNA Rat 159ctctcaccat cgttgtgctt cgtgatcaca accactttct ctgccaccag gtaggcagag 60tagaagccga caactctccc 80 160 80 DNA Rat 160 ctctcaccag aacgaccggttctcgccctt gtagttcacc tgcaccttgg gcttgtcgcc 60 gtcgttcacc acactctccc 80161 80 DNA Rat 161 ctctcaccgg agttggtggt aatttcgatg gccagctgaacgctgcgctg aagggcatcc 60 cgggtcctct ggactctccc 80 162 80 DNA Rat 162ctctcacctg gggcacatct gtcttcaggc ttcttcgagg tcagatccag gctggggcaa 60actgtgactg atactctccc 80 163 80 DNA Rat 163 ctctcaccca tctcagttgtgtttggagaa gtattccttg ctgtcatcca cattgggttt 60 gatggtgtca ctactctccc 80164 80 DNA Rat 164 ctctcaccgt ccgctcaggg agtacaggag gccgataccgagaaagggaa cgatgttttc 60 aaggtcattc agactctccc 80 165 80 DNA Rat 165ctctcacccc cacagccagc agggcactca tgagtatccc taggtctgga ctttgtggtg 60ctactttggg ttactctccc 80 166 80 DNA Rat 166 ctctcaccaa gactgggcagccaccaccta caggatgcat cagctccgtg atagctacca 60 cgtcagccag ggactctccc 80167 80 DNA Rat 167 ctctcacctg agggtaccat atttgcatcg aaggtcctccaccccatcat tcaccatatc 60 caccaaggca gcactctccc 80 168 80 DNA Rat 168ctctcaccca gagctcaaca tgtttccagt gggatggttc ctctgctgcc tccttggccc 60tgagctcaga ccactctccc 80 169 80 DNA Rat 169 ctctcaccag gtgttgggcttgcgctgtgg ccattccggc cttctctgct gcagacaaaa 60 tgttctggga ctactctccc 80170 80 DNA Rat 170 ctctcaccgg gggttgtctg cacatagctg gctagaagaagtggaacgag atagaagact 60 cttagctgta taactctccc 80 171 80 DNA Rat 171ctctcacccg cttcaccacg cagccgttct tatcaatgag aaacttggta aagttccatt 60tgatggcatt tcactctccc 80 172 80 DNA Rat 172 ctctcaccct gtcagttgaggccgcccctc aaacagatta cagccaagag ccaccggctg 60 tatcaactcc tcactctccc 80173 80 DNA Rat 173 ctctcaccag ccactttaaa gcacgctgtc actagaacttcaggacggtc ctgaccacgt 60 caacatagtg gcactctccc 80 174 80 DNA Rat 174ctctcaccgc ctgggtgtgc tccatgggcc cgaactcagc cacaatgtca tagaatgtgt 60tttgcacatc ttactctccc 80 175 80 DNA Rat 175 ctctcacctc ctttgcaattttgataggaa aattccaaga agcctaatga aacagagcca 60 accataagct ctactctccc 80176 80 DNA Rat 176 ctctcacctc tatgtagggt aaatgaaggt gctacacccccacatgtagt gtcataaacc 60 catgttggaa gaactctccc 80 177 80 DNA Rat 177ctctcaccat caggatcaaa aactgcttga tttcttggac ggcaaagagt cttccaggac 60atattgtcgc gcactctccc 80 178 80 DNA Rat 178 ctctcacctg ggtatttcatgaggatcagg agcccatatc tcagagttgt gctggtggtc 60 tcagttcctg caactctccc 80179 80 DNA Rat 179 ctctcaccaa gtccacagca ttccctgagg tgacattctccacaaagtcc ttgctgctct 60 tcacgaggtt gaactctccc 80 180 80 DNA Rat 180ctctcacccc aaaccaaagt gggtgggcat gggggaactg ttgggcccag gataccacct 60tgtccaggct ccactctccc 80 181 80 DNA Rat 181 ctctcacctg taagcaatggctagctgtta aaattatgag ctcagacagc gcttcaaaca 60 catttctttc tcactctccc 80182 80 DNA Rat 182 ctctcacccg gccaatgatg gaatgctctc ctgagagtgagatcacacga tcttcaatgg 60 acacattggc caactctccc 80 183 80 DNA Rat 183ctctcaccat aaggaaggga atctgggatg tatgtgtaga atgattccgt tttttgcttc 60cttcctgaag agactctccc 80 184 80 DNA Rat 184 ctctcaccgg agcggcatcatctgcttctg ctgttcctgc tgctgttgat tctgctttcc 60 tctgctcctg ctactctccc 80185 80 DNA Rat 185 ctctcacctc aacagaatga aatagtgttt aaggactaggctttaaagct acttgaagat 60 atcttagaaa taactctccc 80 186 80 DNA Rat 186ctctcaccgt aagtgaggtt ggtttgtgtg ggtttggggg gaggggagag agaagctcca 60tctctggcct caactctccc 80 187 80 DNA Rat 187 ctctcacctg aagggggtgaagatcccacc caggtcacac agggcagagc ataccatcat 60 cccaagattc ctactctccc 80188 80 DNA Rat 188 ctctcacctt catcacttag ctcaacttca aattcttctgacaggatctg gcttcctgat 60 gtctgcatgt taactctccc 80 189 80 DNA Rat 189ctctcaccgg acgtgcagaa acactcctcg catgttcagc tgtccagccc tgttgtcttc 60tgattccaga ctactctccc 80 190 80 DNA Rat 190 ctctcaccga aaccatgatttcatatcatc aaccaaagca caaagcagag agtgtactgg 60 aagtaggtga ggactctccc 80191 80 DNA Rat 191 ctctcaccac ctgaacctgg cagcccagcc tggcccttccctctcctccc tgcactccag 60 tgctttcaac tgactctccc 80 192 80 DNA Rat 192ctctcaccgt cgacgcctct tccttttctt tcctcatctc attaatatct ttggtgatgt 60cctcagtgcc ccactctccc 80 193 80 DNA Rat 193 ctctcaccgt ccaggccacagctcctcaga gcccagaata aagtccaaac caaagacctg 60 agccacaagg atactctccc 80194 80 DNA Rat 194 ctctcaccgg ttggcactga aatttgagat ttttaccctacggtggtggc agtagccaca 60 ggtcccatct tgactctccc 80 195 80 DNA Rat 195ctctcaccgc aggttagttc ttggacagtt ccggagtgcg actcatcggt cctgcgaggg 60acatgacgca ggactctccc 80 196 80 DNA Rat 196 ctctcacctg ctatttgaggttattactgc agttctaatg gacaacatta agccaatgtt 60 agaaattgct aaactctccc 80197 80 DNA Rat 197 ctctcaccgc tgctggctta acctcttgcc aggtgctatctcacttgact tatggcagtg 60 gcggcattcc ttactctccc 80 198 80 DNA Rat 198ctctcaccca cctgcactca gctccacact ggccaggctg cagcgccgag gcttgtgctg 60acaagctggc cgactctccc 80 199 80 DNA Rat 199 ctctcaccag accacagaggtgatggcagc catgaatgcc atcccagctg gcatgtataa 60 gtggaggtgg cgactctccc 80200 80 DNA Rat 200 ctctcaccgg aagcgtaccc caatatctag taggtacagcaggtcactcg tgtagtccag 60 cacgaaccag gcactctccc 80 201 80 DNA Rat 201ctctcacccc tggatggtat gcagccagct ggtgaaggtc agagagcagt tctgcacatc 60gaacgggaag ttactctccc 80 202 80 DNA Rat 202 ctctcaccgg tcacagtcaacatctgttgt tctgtccaca ccagctccaa tcaaagagaa 60 gctcttggga gcactctccc 80203 80 DNA Rat 203 ctctcaccca gcaaccactc ctcattccca ctgtcaaagtgactttgctg gtgtcgggca 60 cgcgccatga ggactctccc 80 204 80 DNA Rat 204ctctcaccaa aggctggggt gggggctggt tgtagacacc ctgcaggaag atccaggctg 60tgcccaccac caactctccc 80 205 80 DNA Rat 205 ctctcaccct gctggtccaccatccaaatg tggggctcgt ccacaaagga cacatacttt 60 aacgtcggct tgactctccc 80206 80 DNA Rat 206 ctctcaccca gaagcagggg cagggcagct ctgagcagatcgttagcccc tgtcctctgg 60 acggcttcct ggactctccc 80 207 80 DNA Rat 207ctctcaccag gagcatctca atactgttgg ctgggctctc catgatggat ctttggttca 60ggtctttgta gaactctccc 80 208 80 DNA Rat 208 ctctcaccct actgaggactcaatggcggt gtccctcact tggtaggcct tctcatgcaa 60 gaaaacccac ccactctccc 80209 80 DNA Rat 209 ctctcacctg ggatgaggga ttcaggctag cttcctggagataggtgatc cgagttcaac 60 agaatgctca gtactctccc 80 210 80 DNA Rat 210ctctcacctc ttatggctgt agagcttgtt tttgttcatg aacgttaaca aaatccagtc 60acacaggaag gaactctccc 80 211 80 DNA Rat 211 ctctcaccct tcatagacttaatcttgctg atgatgaatg tcttcatgcc taaggatttt 60 atagataacc caactctccc 80212 80 DNA Rat 212 ctctcaccac gtcgcataga gctagggtct gttaccaccagcgattgctg ttcctggaat 60 tgtccatcat ccactctccc 80 213 80 DNA Rat 213ctctcaccct tgagagtgct gcggtaggtt tcgtgcctgg tgcctccgac attgaggatc 60accctctcgt tgactctccc 80 214 80 DNA Rat 214 ctctcaccaa ggtggggagggcacccacat ctgtgcccac ctccacccag tgctttcttc 60 tctgtggcca ctactctccc 80215 80 DNA Rat 215 ctctcaccag tatggaaatc atcgtatgta ccgacacctgggatgggctc atccaaaaga 60 tccagcaagt gtactctccc 80 216 80 DNA Rat 216ctctcacctc cggatggagt tgctcctcct catggagttg tttcttctca tagagttgcg 60cttcctcaga gaactctccc 80 217 80 DNA Rat 217 ctctcacctg agggctcagacctcatggca tcttttagac caggcaatga tcatctagag 60 cctcttgctt gcactctccc 80218 80 DNA Rat 218 ctctcaccag aaaaagaggt gcctatggtc tgcttctgtaagaaacacaa atatcttgat 60 ataattgact atactctccc 80 219 80 DNA Rat 219ctctcacctc tagagctttt aattctggca gcactgtcct caggtgaagc ctgactagaa 60aacaggagaa caactctccc 80 220 80 DNA Rat 220 ctctcaccca ctagcgtgtttgtgaccacg ctcattatgc tgttgattct gtccttccgt 60 ccgtaggagg cgactctccc 80221 80 DNA Rat 221 ctctcaccgt tggtggtgga gaggatgaac atggagctgtagggcggcat gggctttggg 60 ccatcctccc cgactctccc 80 222 80 DNA Rat 222ctctcaccgg ccaggcgaca acctattaaa ctataaggag ggctgggtct gggtctgggg 60gaacccaagg gaactctccc 80 223 80 DNA Rat 223 ctctcaccct ggcgaaagcctggatgctca ccggctggac accacccacc tcctcctcct 60 cggtcttcaa ttactctccc 80224 80 DNA Rat 224 ctctcaccgc tctctggaac tccgttctca ctgtcctcttcctctttgct tgttagggca 60 acttcatttt caactctccc 80 225 80 DNA Rat 225ctctcacctg gggatatgaa gaacaaattt tcattgccag cgtcgacgac aaagttgatg 60ttggtctgat ccactctccc 80 226 80 DNA Rat 226 ctctcaccct cagtccattgggttcctcct cctcatcgtg ctcagcctct gcttctttct 60 cagcctcagc gcactctccc 80227 80 DNA Rat 227 ctctcaccag aagccgaaga aacccagcac catgaggatatagagagcct ccagcttgct 60 gtcatctcgg agactctccc 80 228 80 DNA Rat 228ctctcaccaa gactacggtt ttagtaaagt atctacagag tgggacagga gtcgccaagg 60gttttgtttg ggactctccc 80 229 80 DNA Rat 229 ctctcaccgt attcaaagaaagaaggcagg gaggcgggaa cgggcgattc ttgatagaga 60 gcttccaacc tgactctccc 80230 80 DNA Rat 230 ctctcacctt gatgctgcat ttttttcatc atcatcatcatcatcatcca cgaacattta 60 ttgagcgcct acactctccc 80 231 80 DNA Rat 231ctctcacccg taccgtgtgg atgcacagac acacaagacg acccactatg gacacacgtg 60tgacgcaaag atactctccc 80 232 80 DNA Rat 232 ctctcaccgg tcggtggggaacttcatctg caagtgagag gcagagggtg gtggcagggg 60 tgacctctca gtactctccc 80233 80 DNA Rat 233 ctctcaccat cactgtttct tttgcagtca caaactggaccagcacacgg atccctgatt 60 gaagttttgg taactctccc 80 234 80 DNA Rat 234ctctcaccgt ctgggggagc tgcctccgcc cacctcggcc cggcgtgcta gcaccagatg 60ttgacatcaa ggactctccc 80 235 80 DNA Rat 235 ctctcaccct cctcacacatttgaccacca tcagcaccaa gatgaccaca gccaggaaac 60 cccccactga ggactctccc 80236 80 DNA Rat 236 ctctcacctg gaacccacgt tcctgtgcca gccctggctgcgtgccaacc gtcggaacac 60 ttcctggatg gcactctccc 80 237 80 DNA Rat 237ctctcaccga gagcaccaaa acggcaagct gaaatcttcc atagtccaga gtctgccagt 60gaccacgtga ccactctccc 80 238 80 DNA Rat 238 ctctcaccgc tggtgtcactgtaggtgctg tcccggggtg aagtctcctc agacttgcag 60 taaatgggtg acactctccc 80239 80 DNA Rat 239 ctctcaccct ttcaggagct ccatcttcac cctcaggcacattccagctc tctgcaggag 60 ggggtggccc ccactctccc 80 240 80 DNA Rat 240ctctcaccgt cgactccaga ggtggactgc cacagaagag gcttctgagg gcaatgcctg 60ctgattccat gtactctccc 80 241 80 DNA Rat 241 ctctcaccat gttactgagtctgtgaccca aggaattcgg cctgctctgc tcaaacacag 60 gtccccaagg gaactctccc 80242 80 DNA Rat 242 ctctcacctg catgcgctcc agatccaggt ggctgccaaactcaagcatc cggataatgc 60 ctgtctcctc ttactctccc 80 243 80 DNA Rat 243ctctcaccgc gttaccagga ttatctggtc attgacgagt ttgaggtctt taaacgccat 60cgtcatccgc ccactctccc 80 244 80 DNA Rat 244 ctctcacccg gctgatgtgtcttggcaggt catccctggc ctgcttaggc agctttggct 60 ggtgaatggc caactctccc 80245 80 DNA Rat 245 ctctcaccag gggccctcgc gacgaggcga gggtcagggcatccagcagg ctgcggtcct 60 catcaagctg gcactctccc 80 246 80 DNA Rat 246ctctcaccag gctgacgacg ctcttctcag gggtgttaaa gaaggctgcg gccattgagt 60tgtattcgcc atactctccc 80 247 80 DNA Rat 247 ctctcaccga ggcagtggaagagaggaggg aggtaaagag ggagggagga agggagtggg 60 gcggcgctca gcactctccc 80248 80 DNA Rat 248 ctctcaccta tggttaacac cacaatgagc acgtacatcatgatctctga cacgatggat 60 gccatatctc tgactctccc 80 249 80 DNA Rat 249ctctcaccaa caaggatctc cttcccgttt accgcaaccc atggctcatg tcacgaagag 60tgagttcctg gtactctccc 80 250 80 DNA Rat 250 ctctcaccta ttgacttgggcatgaactca caacttcatt ggaatctgtg tctaatgttt 60 tcaaaatgct ttactctccc 80251 80 DNA Rat 251 ctctcaccat tttcataact gcattcttta tgtatgaatatatcggagct gggcgggcgg 60 cacgggcagg tgactctccc 80 252 80 DNA Rat 252ctctcaccag tacccctgcg ttggtagaag aaacccctcc accccacccc aattaggacc 60tccgcttaca gcactctccc 80 253 80 DNA Rat 253 ctctcacctg cctgcactcacaccctcctg ctgtttagag gggtatccat tctggctgga 60 agacccctgc taactctccc 80254 80 DNA Rat 254 ctctcaccct accgagaaaa tatatatggc tagcaagacatatatgtttt ctcttaggtg 60 gtttatgtat tcactctccc 80 255 80 DNA Rat 255ctctcaccta tacttgcaga gaatactaag gttgactctc tctcagggag acacatggtg 60ccacttatat agactctccc 80 256 80 DNA Rat 256 ctctcaccgg aaggtgatgatcatgcctcc cttgaggatg tagggtacat agtgtggcct 60 tttggacttg agactctccc 80257 80 DNA Rat 257 ctctcacccc caggatcagc acaaccactg tgacccacactgggtctcca gaatgaagcg 60 gagttgtcca ccactctccc 80 258 80 DNA Rat 258ctctcaccgg taagagggat acccagcaag aagccagcca cagccacacc caaggtcacg 60taggtcttct tcactctccc 80 259 80 DNA Rat 259 ctctcaccgt ctccagagctgttctgcagt ggcgctggcg actgtgtttc tttcattgtc 60 ttcttgggct tcactctccc 80260 80 DNA Rat 260 ctctcaccag gcgggtagat gcacgttgat tgtatttcaaaagatatgaa ctctgtctta 60 tgaggttgac agactctccc 80 261 80 DNA Rat 261ctctcacctt gcagtgtcca cctggaaccc tgctggtcaa acaaggccca aaacatgggg 60aggggaatgt acactctccc 80 262 80 DNA Rat 262 ctctcaccgg cctcattcatggtctcattc aacagggaga tgactgcctt ggttgtattg 60 gcctcctcgg atactctccc 80263 80 DNA Rat 263 ctctcaccaa acgaagacca gcaggggtgt ccagagacctgggaggtctc aacagggtgg 60 aagagaggtt caactctccc 80 264 80 DNA Rat 264ctctcaccac cctagagttt ctgctttagg gcttgaacca cagtaaccca gcaacttgat 60ttctcacccc agactctccc 80 265 80 DNA Rat 265 ctctcaccat aaatcctgtgcagacgagca agcccctaac attaggtgaa atacaaggtt 60 caggattcat ttactctccc 80266 80 DNA Rat 266 ctctcaccat acaacatgcc tgttctcggg gagaacacctaggacataat tatgtgtact 60 tcttgatttc atactctccc 80 267 80 DNA Rat 267ctctcaccta gtaggccatg acggagaaga tcaggcagac caccaggagg aggcaggaaa 60acaaaacgaa ctactctccc 80 268 80 DNA Rat 268 ctctcaccct ggccagtgctgccaggatgg ctgtagtggt ggtaccaatg ttggagccca 60 gggtgagagg gtactctccc 80269 80 DNA Rat 269 ctctcaccta cagctcccac tcggggaaag tcctgggttggtgaccagaa aaggcaattg 60 tgcttgtggc ttactctccc 80 270 80 DNA Rat 270ctctcacccc tctgatacag gagggcccag ttaaggccaa ggaaggttca gctccctcct 60ttcccggtag gaactctccc 80 271 80 DNA Rat 271 ctctcacctg atagatgcctctcagaacac cccgggtaca gtagccaagc acagcctgcc 60 ttggaagtac ccactctccc 80272 80 DNA Rat 272 ctctcaccgc gaggagtatc cagagggtga cttcataagggatttgaaca tagtcataat 60 ccaattcaaa aaactctccc 80 273 80 DNA Rat 273ctctcaccca aaggcctcgt ccctagcagt tagactccag ttctgtgagc ctgaggaggg 60acgtcatcgg tgactctccc 80 274 80 DNA Rat 274 ctctcaccca ttgccctgatgctgataaag tcaggatgag agtcagaaga aagggcaaac 60 ctctcagcta ccactctccc 80275 80 DNA Rat 275 ctctcaccgc aggatcaaga atatacccct ctttgcccctgttggggtaa acaggaacat 60 tcaaagactc caactctccc 80 276 80 DNA Rat 276ctctcaccac tatcttgacc tgaagagtct tcatggtctc gtcatcgccg aactccagct 60ctccacaagc atactctccc 80 277 80 DNA Rat 277 ctctcaccgt gatttcctctaaccagtagc acaggaggga ggttcttggt caggatttct 60 aaccacgcca tgactctccc 80278 80 DNA Rat 278 ctctcacccg cggatcttgc atttgcccca tctcttcttgcggtgcagga gataagggat 60 gaggagggtg agactctccc 80 279 80 DNA Rat 279ctctcacctc cttctcactg ctgctcatat atgggtggtt gttggggtca tcaaagagaa 60ggatgaacca gaactctccc 80 280 80 DNA Rat 280 ctctcacctg atgtcctctagccgcctggt ggttgcagcc acctcctcgg tagtgggtgg 60 aggactccct gaactctccc 80281 80 DNA Rat 281 ctctcaccgg aattagacgg attgagaagg cactggcttgtgttggagtg ggcccaggag 60 gggaatgtgt ccactctccc 80 282 80 DNA Rat 282ctctcaccac ctcagacaca aacattggag aggtgttccc tgtgtttccc ctgctgccct 60tggtcagccc agactctccc 80 283 80 DNA Rat 283 ctctcaccct cactgcttctcatagtagaa ccaatgctca ttgataaaaa tgaaccaagc 60 tcaacccttt gcactctccc 80284 80 DNA Rat 284 ctctcaccca tgagggttgc tctggagtga aagggcgtagccccttcacg ctccacagcc 60 cagcggttgg gcactctccc 80 285 80 DNA Rat 285ctctcacccg ccaatgatca cagcgctgac tgtgagcaag atgaaggcat ttcggcgcag 60gaagcgccgc acactctccc 80 286 80 DNA Rat 286 ctctcaccgg ataaaagatcccaccagcat gatcgcagtc tgcaaggtgt ctgtgtaaat 60 tactgctgcc agactctccc 80287 80 DNA Rat 287 ctctcacctt ataaggaaag aaacttctaa actactaactaaaaaaaaaa aaaaaatcaa 60 cacaatagca tcactctccc 80 288 80 DNA Rat 288ctctcaccga ccaggttgag tgcgtccaca aagcaggggt tttggagaac accatggtag 60atgttgtagg agactctccc 80 289 80 DNA Rat 289 ctctcacctg ctggggtacaagattcttga agctggcgat gatatttgtg ctcttgaact 60 cctggtagta gcactctccc 80290 80 DNA Rat 290 ctctcaccgc cagcaggggc tgcaggtact tgggctgcaggagtttgccg tagtagggat 60 agtactgcag agactctccc 80 291 80 DNA Rat 291ctctcaccaa gtaaggccgg tgacgacatc gttctcgtta tcccactctg gttctggctc 60tcctgcgggg aaactctccc 80 292 80 DNA Rat 292 ctctcacccc gtgaggaaaccatagaagac gaggtagaag aggaggatga aggcccagct 60 ggtcccggtg cgactctccc 80293 80 DNA Rat 293 ctctcaccga caggcagaga catctgaagc accaccccccactgtcgacc actcagtggg 60 gtcacctgga aaactctccc 80 294 80 DNA Rat 294ctctcaccaa cttggtatgg ttcggagcag aaaaggtctc ctggaagaag tgcttttcag 60acgaacagtt gaactctccc 80 295 80 DNA Rat 295 ctctcacctg ttcttatcccaccagcttcc agggtagagc ctgatgaaca actttcgcat 60 ctcgtcatac acactctccc 80296 80 DNA Rat 296 ctctcaccca gcaccatcag ccactgggtc agattcagcggtgtgatctg gaaaatgagt 60 ggcaaaggtt ccactctccc 80 297 80 DNA Rat 297ctctcaccag agagcacgag gagaagtggc agctaatggg gctctcagaa cagcacgagg 60ccccagagaa ctactctccc 80 298 80 DNA Rat 298 ctctcacccc gtggtgcctggctggggaag tggcatcact gtgcccatgc accggtggca 60 ctgccggctc ttactctccc 80299 80 DNA Rat 299 ctctcaccgc aatcccaccc agcctctgca gatgcaccacacactgctgg cgaggctagc 60 agggtgcttg ccactctccc 80 300 80 DNA Rat 300ctctcaccgc aactgctcca gctttcgcat aagcctggag ttccttatta gtaaatgaag 60tcagtacctt tgactctccc 80 301 80 DNA Rat 301 ctctcaccgc ttacacagggcattcaggaa gccgaaggcc gtcgtcctct tgtcggaagg 60 gtagagttca acactctccc 80302 80 DNA Rat 302 ctctcaccca cattctcatt cagaaggtct ctgtcttcactgggacagtc cgaagggaca 60 gtggtgttag ccactctccc 80 303 80 DNA Rat 303ctctcaccct ctctcccttc cccctggggt tcccttgagc tgctagtttg ttttgaaaac 60ttggtccctt gcactctccc 80 304 80 DNA Rat 304 ctctcaccgt cctctgtgaacggccctcca cgcgacatct tacatgaatc aaacagactg 60 tgtttagaag atactctccc 80305 80 DNA Rat 305 ctctcaccca tcagttaaca ttttggtaat attcacatagtttgtgttag ccaaggtaca 60 gttggctgtt ttactctccc 80 306 80 DNA Rat 306ctctcaccgg gttccaaacc caacacagta aaaaagaaag agggctggaa aaatgaacag 60tgtgcattag caactctccc 80 307 80 DNA Rat 307 ctctcaccag cccacctcagatgatcttgc agtcttcctt gagcattaag gcagaaaaca 60 gagctggcct agactctccc 80308 80 DNA Rat 308 ctctcaccag cagcggcacg tcgtagctca tgcgatcaatgaaaggcccg cttaaggggt 60 tcaccagaag ctactctccc 80 309 80 DNA Rat 309ctctcaccct catccacgaa gccccgccac agcgtcatgt gttcaggaag ataggtggca 60aaaatgtttg caactctccc 80 310 80 DNA Rat 310 ctctcaccct cttctcctcctctggatcca gcttcctgga ggatctcctc gaaccgctcc 60 acacctaagg taactctccc 80311 80 DNA Rat 311 ctctcacctc agccaaatat tggactaaaa ctaaatcaaaacatgaaata atgcctaatg 60 gggtgttttg ctactctccc 80 312 80 DNA Rat 312ctctcaccgg aactgggcgt atggttgcgc ctacatttgc acttgccact cagggccatg 60gcaattccag cgactctccc 80 313 80 DNA Rat 313 ctctcaccgg cgacgtttgagctccacgtc aggacagaag acatactcgt aaagtgcacc 60 tgccagcaca gcactctccc 80314 80 DNA Rat 314 ctctcaccag ccacgcgatc atggaggctc agagaggaggggaagagcag gtagaaatag 60 aggatggccg ccactctccc 80 315 80 DNA Rat 315ctctcaccgg ctggcacggg atgcagaggc cttgggagtc cctcagtggc tccatcgtgt 60cctcgttgac ctactctccc 80 316 80 DNA Rat 316 ctctcacctg ccccagacaaggctagacca tgtgggacag ccaggtcttg gggagtgact 60 ttggtcagct tgactctccc 80317 80 DNA Rat 317 ctctcaccag caaccggtga aatagatccc aaggaggtggcccagggtaa cagagaaacc 60 aatggagagg gcactctccc 80 318 80 DNA Rat 318ctctcaccgc ttaatgagaa tgtatctttt cagctctaaa tacttccaac tgtgaaccat 60aaattcaatg aaactctccc 80 319 80 DNA Rat 319 ctctcacctg cctggtgggcaggctgaaag ccctgagtct ggaagttgat gaaggacagg 60 tgcaagcatc ccactctccc 80320 80 DNA Rat 320 ctctcaccac aggagcggat cagaactgga gaaatgagcagtgccttcgt ggtctgcatt 60 tttcaatctg caactctccc 80 321 80 DNA Rat 321ctctcaccga gaggtgctcc tgatcaggga gcgggtagag acgaacttgg agcaggcata 60catttttcag ggactctccc 80 322 80 DNA Rat 322 ctctcaccat tccatgggagaacaccatac attctggcat cagcgcagag actgcctata 60 ctggcagagg tcactctccc 80323 80 DNA Rat 323 ctctcaccta gttcttgggg tagggactgt tgaaataccagacgccatgt accacagccg 60 gtctgagctc atactctccc 80 324 80 DNA Rat 324ctctcaccct atcctggtga gctggctaat cttatcattg ccaagccaat attcgcctgg 60caagccacag taactctccc 80 325 80 DNA Rat 325 ctctcacctc ctaaaacttagagccaacaa tacttctaag gtaagaatat atttcccaca 60 ggtagcactg acactctccc 80326 80 DNA Rat 326 ctctcaccga tgatgacagc attgtctcca tcgtcgcttttcctacacgc cgtgaggtcc 60 ggcagccggt ccactctccc 80 327 80 DNA Rat 327ctctcaccat ggggtaaagg gaagtggcca tgagagaagg cagcacggac agtggtcagt 60cactgggcag gaactctccc 80 328 80 DNA Rat 328 ctctcaccca gtgaatactgcagtcggcct aactggtgct tgtccaatac ctgctgtcct 60 ggcatggatg gtactctccc 80329 80 DNA Rat 329 ctctcaccac ccttcagttc cagatccagg gctttgttcatgtcatgttg actgtacttt 60 ctataattct gaactctccc 80 330 80 DNA Rat 330ctctcaccgc cacatggaca tcttatttca tcccctggtc ccctgacctg caagcagctt 60tcaggacgcg ctactctccc 80 331 80 DNA Rat 331 ctctcacctg ataacctcaacctccaactg acccttctta tcctccatcc cgatttggat 60 atcgcccatg gcactctccc 80332 80 DNA Rat 332 ctctcaccag tagtacaggg atttgccata tttcctcttgaattcagatc tgattttcaa 60 catgtccact tcactctccc 80 333 80 DNA Rat 333ctctcaccag tgcaggccct gcccagggca gctgccagaa gaggacccaa gccctgtccg 60gtggcgcggc caactctccc 80 334 80 DNA Rat 334 ctctcacccc tttcaaggaagggagaatct gggcaatgga catggggttc tggaatttcc 60 cctgcttaat ctactctccc 80335 80 DNA Rat 335 ctctcaccag gggtgaagcc tcctctgggg gtgatggtgtctttgcaatg cgaagaggac 60 ccgaatcact ggactctccc 80 336 80 DNA Rat 336ctctcaccgc tgaaggatgc ggaagagcgg gtagagatgt atgggtagtg ctttgtatcc 60agtttgtctt caactctccc 80 337 80 DNA Rat 337 ctctcaccag gcaaagctgctttacgcttt gcttcactct tgtcatctcc tccaaacttc 60 tttttgctac ttactctccc 80338 80 DNA Rat 338 ctctcaccca atgagtggca tggaggagcc caccacctcaataatgtgat cccttccgtc 60 cttgccatgc agactctccc 80 339 80 DNA Rat 339ctctcacctc ctgagagcct gttatcctca aagcccctca ttccctgaaa ttatttggtg 60ccttcatgaa caactctccc 80 340 80 DNA Rat 340 ctctcacctt ctgttttgtcctactcctcc ccaaggcagt gatgaccccc acacacacta 60 gagccacccc tcactctccc 80341 80 DNA Rat 341 ctctcaccaa ataaaaccaa agaagtgaag cagtgtggttctgtacgacc tgctcattga 60 attgagctat tcactctccc 80 342 80 DNA Rat 342ctctcaccag ttcgcactcc tcccccaagg tgaactcatt gtggatcacc ttggacccat 60aggtgatggt gaactctccc 80 343 80 DNA Rat 343 ctctcaccac acaatgggacctgggtccag gctggtgtcc catatctcca ggaaaagccc 60 aaccaaagag taactctccc 80344 80 DNA Rat 344 ctctcaccct ccttcatatg tgtaggtttg gattagttcgttaccagaaa tctctcggac 60 agcaatcagc tcactctccc 80 345 80 DNA Rat 345ctctcacctg tcctgaggtt gccaaaggga ctcttcacag cttagctttg tccggctgaa 60gctgcaggct ttactctccc 80 346 80 DNA Rat 346 ctctcaccat cccacttctgcacatgtacc aggaccccac catccagggt tatgatgctc 60 ttcactttcc tgactctccc 80347 80 DNA Rat 347 ctctcaccgg tcctagtgct gacaacattg ttcatggtgcactccactac cattcttcca 60 tccacaattc ttactctccc 80 348 80 DNA Rat 348ctctcaccca atatcactac ggagtaagtt gagtgctatc cactgaggaa actacgtgtc 60taagcacacg ggactctccc 80 349 80 DNA Rat 349 ctctcaccct ctaataggtctctggctctg tgcaaagcac ttatcgtcag atggtatttt 60 tccacttaca caactctccc 80350 80 DNA Rat 350 ctctcaccgg gcagtgcagg tggtcgctcc tcactgcttgggtctgcttg ggtggtcagg 60 gtctgctggg tgactctccc 80 351 80 DNA Rat 351ctctcacctc cacatgctct gagatcttgt cgatgttgag cctcgtttcc tcaatctcag 60aaaagaactc atactctccc 80 352 80 DNA Rat 352 ctctcaccta cgattgatcatctctccctg catctccacc tcggtagcta gaaaagtgaa 60 gatctcatgg agactctccc 80353 80 DNA Rat 353 ctctcaccaa gaagacaatg aggatgagga agattttgaccatgagccac cgattggagg 60 tgactgactg gaactctccc 80 354 80 DNA Rat 354ctctcaccgg gagttggcca gctgcagctc ccgatcaagg cgcccatatc gatccgtcac 60tccagcatcc caactctccc 80 355 80 DNA Rat 355 ctctcaccct gacatctccaaagaagccaa ggtgggggca ggaggtgctg ggccctgcag 60 cagaatattc tgactctccc 80356 80 DNA Rat 356 ctctcacctc tgcaggaagc agcagagagg agcataaatgatgttgatgg taccaatgat 60 gaccatgagc caactctccc 80 357 80 DNA Rat 357ctctcaccag tagctggata actctggaac ttcaggaaac caagttttca aaagacccgg 60catttcttct atactctccc 80 358 80 DNA Rat 358 ctctcaccga tcttgtcatagggtggcacg gaggcatcca acagcaccgg gacaatcagt 60 ttcatggtgt ccactctccc 80359 80 DNA Rat 359 ctctcaccat gacggaggtt tccgatgatg ccgctcacctgctccaggtt ttcatccatt 60 tcattttccc ggactctccc 80 360 80 DNA Rat 360ctctcaccaa ccaggaagat cttggtagtg ccccctttga cgcaggaggg tgcatcaaag 60tacacttggt gcactctccc 80 361 80 DNA Rat 361 ctctcaccct gcaggtgggaagtggcggcg gcacacaggg cacgttccgg acttctgtag 60 ccaaatggag acactctccc 80362 80 DNA Rat 362 ctctcaccct ccacctgact gcttgactgg agtgctctggattttaacat tggacttctc 60 cgcacctcca gcactctccc 80 363 80 DNA Rat 363ctctcaccgc ccaaggagca cttgttgctg ttcttcttgt cagatacttt caggtcagtg 60cttgagggct tgactctccc 80 364 80 DNA Rat 364 ctctcaccat atcttgccaaagactcgctg taaaagttta attctgttaa tcgttcacaa 60 catggctggt ctactctccc 80365 80 DNA Rat 365 ctctcacctc aatctggcaa agccctgagg cagctcacccaggctataga gtgggtacaa 60 atatgggctc ttactctccc 80 366 80 DNA Rat 366ctctcaccgc gatgtctcct gcaactacca aagcgtatcg tccatcccaa gagctggatt 60cgatccagtt caactctccc 80 367 80 DNA Rat 367 ctctcaccgg gtccaggcgagagatcagat caacagcaca cgcgctctgg gtggtattct 60 acgtggtgct ccactctccc 80368 80 DNA Rat 368 ctctcaccgg catcacctcc aaatagtctc cacgacgcacgtgaacaccc acataggtac 60 gaggacggat gcactctccc 80 369 80 DNA Rat 369ctctcaccgt gggaccgagg acaggctggt gttaatgcct taaggagtgg ggacagatcg 60gcagggatgc ccactctccc 80 370 80 DNA Rat 370 ctctcaccag ccaatgcactggcctggagg gctctgccgt atgaaaagct tagtttccag 60 ggcctaggta gaactctccc 80371 80 DNA Rat 371 ctctcaccga gagattcact ggctgcggct ccagactggccacttggagt gtactttcct 60 tgacaagcga ggactctccc 80 372 80 DNA Rat 372ctctcaccag gattcttcct catgttaaat cttgatggtg gagtacagtt catctatctg 60tgacctgaaa taactctccc 80 373 80 DNA Rat 373 ctctcacccc gtcactggttgtcaccaagg aggcaggtgg gttaggctga gggttctgag 60 tgttaccagc caactctccc 80374 80 DNA Rat 374 ctctcaccaa atccatagga attaagttta tatccaggaggttgatctct tgtgaacatt 60 ccccaagtgt gcactctccc 80 375 80 DNA Rat 375ctctcaccaa agttcgtcac agatagaatg gaccggtgac tctcgagctc tgctctgggg 60caggggcagg gaactctccc 80 376 80 DNA Rat 376 ctctcaccaa gaacagccagagcaggtgag tcgaggtgtt ctgggtgact tggggctcaa 60 ggtatcaagg taactctccc 80377 80 DNA Rat 377 ctctcaccct agattggggt cttccttggt gcccgaagctgcctttggag gtagcatatt 60 gtatggatcc agactctccc 80 378 80 DNA Rat 378ctctcaccta ggacttcttc cccaccagga tgctcttcga gaaacttggt cagatcgtac 60accttatgat gtactctccc 80 379 80 DNA Rat 379 ctctcaccat acttcttcttctgttgatca tttccagcaa taatcacagg catttgcccc 60 aaagaatttg ctactctccc 80380 80 DNA Rat 380 ctctcaccgt tgtgatgtgg ctggcgcttg attcggtaggcggaggagtc agaaaccggg 60 aaggcattca tgactctccc 80 381 80 DNA Rat 381ctctcaccaa ctcatgagga tgttgcctct tttaaatatt gaataaggtc ttctctctca 60ctcttctttt tgactctccc 80 382 80 DNA Rat 382 ctctcaccca caggggggctctggtccaca gcagaacctg tggtgctcac tgcttcaggg 60 agggtcaccc agactctccc 80383 80 DNA Rat 383 ctctcaccct cagtgaagag gaagggaggg gctggaaaccaaggcttgaa acttaaatag 60 agggtggcaa acactctccc 80 384 80 DNA Rat 384ctctcaccgg gagggaggag cccacagcag caggctctca cttcttccat tcattcttgt 60tgtagtccca ctactctccc 80 385 80 DNA Rat 385 ctctcaccgg agtggagattcccaattcat ttaaagttgg tctaagttcc tggatgacat 60 aggggtagat ttactctccc 80386 80 DNA Rat 386 ctctcaccga atgcagccat agctaacata tagatggcatacgctgttcc accaactgtc 60 agaagcattg tgactctccc 80 387 80 DNA Rat 387ctctcaccct tctcttatag ctctccaagt gggataagac ccatcctgct ggaaccatga 60agccagcgac taactctccc 80 388 80 DNA Rat 388 ctctcaccga ggttctggaccatctctttg cccaggtagt ggtcaatgcg gtagatctgg 60 tcctcacgaa acactctccc 80389 80 DNA Rat 389 ctctcaccgg gacatccgag ggcacaggag acataccctctcacagagat gctggcagcc 60 ctcgcggcct gcactctccc 80 390 80 DNA Rat 390ctctcacctc tcatattccc ggaccctagc ctgagatcca gtcacgaact gggcacagtt 60cttcacatcc tcactctccc 80 391 80 DNA Rat 391 ctctcaccag tgctgttgtgtttggctgaa gggcccctca cgacccatgc tcctctgcca 60 gcttgtcagc ctactctccc 80392 80 DNA Rat 392 ctctcaccat cacatgcttc tccacccagt ttatcatgtgctctccctcc ttgcgacgca 60 tcatgtcctg taactctccc 80 393 80 DNA Rat 393ctctcaccca gtcaaagagc caccaccaaa ggaatcgttc atcttggctg ctctctccag 60caggcgagag tgactctccc 80 394 80 DNA Rat 394 ctctcaccga cccaggacgggacaggatgg gagggaggaa gacatctagg gctgaggctc 60 tgtatggccc tgactctccc 80395 80 DNA Rat 395 ctctcaccta gcatggcttg gggtacttga cagggcagttccacctcctg ttccaacatc 60 gactacatct tcactctccc 80 396 80 DNA Rat 396ctctcaccac ggagcagaat cccggcggcg agagacccag ctccagccaa agccaggagc 60ttgagtccgg ggactctccc 80 397 80 DNA Rat 397 ctctcaccct gctcagtagagattgtgcca gacgtctggt gtctatggaa agggcagaga 60 ggtcccctgt tgactctccc 80398 80 DNA Rat 398 ctctcacccg agagttagca aagctggtgt catccagctgagggaatctg gcctggaggt 60 aagctcgctt ctactctccc 80 399 80 DNA Rat 399ctctcacccg cacagtgtcg ctgctacaga agttgtactc tttcccgtct acaccacttt 60gcactgtgat ctactctccc 80 400 80 DNA Rat 400 ctctcacccc atgtcagactctgtaacaca atcaagaaat gtcttccact cttgggaggg 60 ctgctgtatg gcactctccc 80401 80 DNA Rat 401 ctctcaccca tagaccctga ctgaaggtgt ccatgttgctgcaatggaag cctgtgttct 60 gcaggttgct gtactctccc 80 402 80 DNA Rat 402ctctcaccac tttgggtaga gttgggttga tcacattgtt gtaccaaata aatttaactt 60tctgcaaatc tcactctccc 80 403 80 DNA Rat 403 ctctcaccga tatcatatatatacgttgtt cccaaggact gtgcctgaaa cctctttgat 60 tgaagaaggt ctactctccc 80404 80 DNA Rat 404 ctctcaccgc aagccacact ctcccgctgt tgtccagctatgcagccttt gactaccaag 60 cccctccctc tgactctccc 80 405 80 DNA Rat 405ctctcacctt tccagcagcc tccgggcact gaggaggtgg agctgatcct gctagaacca 60gcctctcttt ctactctccc 80 406 80 DNA Rat 406 ctctcaccaa gcacacatttcaaagcaggt aaaaagccat tatgaacatt cacccggccc 60 ttcagtcgcc acactctccc 80407 80 DNA Rat 407 ctctcaccta tcagctcttg ttttcaaagc aaaagagtcgctaccagctc cagcttcaga 60 gaggcaaaaa ctactctccc 80 408 80 DNA Rat 408ctctcacctc ctgggcccag gactcagtgt agatgtcaaa ggtggcttgg gtgcctttaa 60gccccttggc gaactctccc 80 409 80 DNA Rat 409 ctctcaccga gccgggatcttttcctccag gtggtccagg cctctgcagg ccaactcatt 60 agcagctgtg aaactctccc 80410 80 DNA Rat 410 ctctcaccgt tgattcccat ccccagaata aacaggaagacaccaaagct aaaccgcaca 60 tctgtgtacc acactctccc 80 411 80 DNA Rat 411ctctcaccga ttggagtcag tgtgttgtat tattggcctc tgagagtcga gcatcacaca 60aaggagcttc gaactctccc 80 412 80 DNA Rat 412 ctctcaccca cattatttacagctaataaa taaaatttaa agtcaaagca tcattgagat 60 atgtacgtat atactctccc 80413 80 DNA Rat 413 ctctcacccg agacaccacc ttctggagcc cttcctccgaccttgcagtc aataccacat 60 gggctcccat ttactctccc 80 414 80 DNA Rat 414ctctcaccga ctcagggtcc acgatctcct ccaacatccc caggcgctcc ccaaatacaa 60cactggtgat ggactctccc 80 415 80 DNA Rat 415 ctctcaccat tttgacgaagattttctcaa agtcagcaaa gtcatgccag gaagactgga 60 acatgtgcat gaactctccc 80416 80 DNA Rat 416 ctctcaccgg gcgggagccc cccagatgat gccgagaacaggaatacacc caggcggcca 60 tgtgatgcca tcactctccc 80 417 80 DNA Rat 417ctctcacctc cgcatgggct tgatggaacc gccccatggg gccccaagag agatgaagcc 60atcaatgaag tgactctccc 80 418 80 DNA Rat 418 ctctcaccga cagaactagggaggctttgg agcaggagag tgatgtggtt acagccagct 60 ctggagggtc tgactctccc 80419 80 DNA Rat 419 ctctcaccca ggcagtttta ttccttctac atgatgtcccctgcaccttg ttcaggtgac 60 tgggccagtg acactctccc 80 420 80 DNA Rat 420ctctcaccag caggcgctcc aacttcagca ggataccaaa ggccagtaga aaacacagca 60tacttaacgc ctactctccc 80 421 80 DNA Rat 421 ctctcaccag tcctcttttctttctttgtt agactctgtg gtcttgaaga aatcagttac 60 atacaaaacc acactctccc 80422 80 DNA Rat 422 ctctcaccgt acatcctcct caagtcccaa tgacttccatgtggtttccc tttcagtgaa 60 ccagatactt caactctccc 80 423 80 DNA Rat 423ctctcaccac gtttcagagc taatgtcatc tctgagattc ttcttatatg ctgtataata 60ggcctgcgag atactctccc 80 424 80 DNA Rat 424 ctctcacctc aggcatgaactgatctggct ggtcccattc attctcatcg tgatgcagtg 60 cccagagatt gaactctccc 80425 80 DNA Rat 425 ctctcaccac cgggacagtc atgacagact gtcacaatgtcacaaacaag catcttcggg 60 tgacacacta tgactctccc 80 426 80 DNA Rat 426ctctcaccat gtactggtat atgatagctt tgacagctgg catgttggtg gcatccatca 60tgagggtaac tgactctccc 80 427 80 DNA Rat 427 ctctcaccag gcaggttgcccagtcgcgcg gctgcaggac gaatgtactc gtacagcggc 60 caggccatcg agactctccc 80428 80 DNA Rat 428 ctctcaccac tctcatgaac tctccataca tcttattgtagtagttgcag gcacttccaa 60 tccccatcca caactctccc 80 429 80 DNA Rat 429ctctcaccgt gtgtctctgg acaggggatt agggaagaaa gcttgtggac tagcaaggct 60tccagtgaag tcactctccc 80 430 80 DNA Rat 430 ctctcaccag taccaagaatgaagtagaca ggaccctcag aagagacatt acagatgaga 60 gcttagagag tcactctccc 80431 80 DNA Rat 431 ctctcacctg gggatttgaa gcagaagttc tgcatgatgtttgtgaggaa gagaaagagt 60 tccatccttg ccactctccc 80 432 80 DNA Rat 432ctctcaccag gggattgcag aaggcaacac agtagaagcc aaaatgcctg atctattcag 60gaaggaatga agactctccc 80 433 80 DNA Rat 433 ctctcaccac aaactgggatggagtctgga ggaaaagcta cagaaatgag ggcaaaaaaa 60 tgagatgacg ggactctccc 80434 80 DNA Rat 434 ctctcaccgc agatggagtg gggacaagag tgactgacagaaagacaaat ggcagacagc 60 atgctgttgg agactctccc 80 435 80 DNA Rat 435ctctcaccac acgaggacag gaggacctgt ttgctcacaa gtagaagcag ggtctcagtc 60atctgtgtga aaactctccc 80 436 80 DNA Rat 436 ctctcacccc tccgtaagattctggtatgg gtgacagttc tgttcattct gggatcctca 60 gaggtaggat ggactctccc 80437 80 DNA Rat 437 ctctcacctt ctgtagaaga tataggtctg agtcaagacacggaccaaga gccaggaagc 60 tccgatcagc agactctccc 80 438 80 DNA Rat 438ctctcaccga aataaacaac ttcagttttc tccagttgct gaaaagtgtt tggtgtgatc 60caggaagata ctactctccc 80 439 80 DNA Rat 439 ctctcaccag tgaacaaaattactgcaatc ttcatctctg gattcattca caatatgaac 60 tggtttctgg tcactctccc 80440 80 DNA Rat 440 ctctcaccac aactgtaaga taataacgtc ctgttattcccgaccatcaa ggttgtgacc 60 gttgattatt ggactctccc 80 441 80 DNA Rat 441ctctcacccc tctacagtgt gctctcccac agaagggtcc ctttgggaag aaggaacaga 60aggggttctg gaactctccc 80 442 80 DNA Rat 442 ctctcacctg gtcaggccagagatttgctg caagttaaag gccagcctgc tgtatccact 60 aaaatttgag acactctccc 80443 80 DNA Rat 443 ctctcacctc ctccacaacc tcctatcttc tttgaaaactttttgtccct ggcttccgac 60 ttgccggaag ctactctccc 80 444 80 DNA Rat 444ctctcaccaa aaacctatga agggtccttc aatgcccctt cttagggagc tggtggcaga 60gttcacgctg acactctccc 80 445 80 DNA Rat 445 ctctcacctg ctgccagagggacaccaaga gcatgacgct cagggaggcc aatatgacca 60 ccagaagcag tcactctccc 80446 80 DNA Rat 446 ctctcaccgc agtggcccaa ggtattctga cacatccactttgaattctg cttccgagtc 60 ccgacagggt cgactctccc 80 447 80 DNA Rat 447ctctcacctt cattggaagg cagagggaag ggggtggggt tggaaggaag gagagtcagg 60ctttcgtcac agactctccc 80 448 80 DNA Rat 448 ctctcaccat ttaccaatccttttagtctt tcaacagtaa gagaaccaaa cattagggga 60 actaaagggt caactctccc 80449 80 DNA Rat 449 ctctcaccaa tgccgcgttt gtcatagaag gagatgacaggttctgtggc tttgtaatac 60 gtctcaagcc tcactctccc 80 450 80 DNA Rat 450ctctcaccga gcccttaata aaccacaaca actcctccaa aacacccttc cagaatactc 60gtttggttgt gaactctccc 80 451 80 DNA Rat 451 ctctcaccgc atacttgatgaccaggcact tgtactgggc gatctggaag cgccttactc 60 tcctcatcag ctactctccc 80452 80 DNA Rat 452 ctctcaccca aagggcacgt tgagatttat cactgtgtctatctgataaa ctctatccag 60 ggcttctgcc tgactctccc 80 453 80 DNA Rat 453ctctcaccgg aggagaggca caggaggcgg atctcagacg gacagtgaca caaagctgtt 60gtactgctgg atactctccc 80 454 80 DNA Rat 454 ctctcacctc cgtacattctaaaaacaaac cacattttca gggagcgaaa tcttcgcccc 60 agtgggattt gcactctccc 80455 80 DNA Rat 455 ctctcaccca tattgggaaa aaacattctt ttttctaaaagaacaagccc ttgggaggag 60 cagcgttggc ctactctccc 80 456 80 DNA Rat 456ctctcacccc ttcctctggg ttgggtctgt gctctggcag tgtgaatcca gggcagccta 60gcctgggaaa tgactctccc 80 457 80 DNA Rat 457 ctctcacctt ttggaagggttcaagtggaa cacttctggt tctaaaaggc cagcctgaag 60 tgtcttcaga aaactctccc 80458 80 DNA Rat 458 ctctcaccct gtgcccactg gtacagctgc tccaactgtgttttgtctag atccgaggtg 60 cctatagcaa caactctccc 80 459 80 DNA Rat 459ctctcacctc catgaggatg taggaggctc tctcctcgct gtctttcagc tgctccagag 60cgtgtaccat ttactctccc 80 460 80 DNA Rat 460 ctctcaccaa gatccagagccgacagcaac cctgtgctat gtatttcttc agtaatgtac 60 agtccttctc tgactctccc 80461 80 DNA Rat 461 ctctcaccct tcccaggtgc caggaagagg ctgaatattattcattagat caaaatgata 60 ataggacaca gcactctccc 80 462 80 DNA Rat 462ctctcaccag cagggtggta ggctcccagc aacggccttt ccctgggtcc ctaggacact 60gcagtggggg taactctccc 80 463 80 DNA Rat 463 ctctcaccta atggaaatcaatcactttag ttgatctatc aaaacttttc accacgtact 60 gaagcaaaat gtactctccc 80464 80 DNA Rat 464 ctctcaccgc agtaaccaca tggagttcca agcccgatgaggtcttcttt acttccttaa 60 cctgtgagaa ctactctccc 80 465 80 DNA Rat 465ctctcaccca ccgtagatcg gctgaactcg agcatccacg taggcttttg caatcgggta 60ttcccacatg taactctccc 80 466 80 DNA Rat 466 ctctcaccgt cattcaggtctctgtggccc tgcctccagc aggtcgctca aagagagagg 60 cactgttggt ggactctccc 80467 80 DNA Rat 467 ctctcaccca atggctgaaa gcctggaggt aagacaagtaacttaactgt gtcaaaaagt 60 ggggtcaaag gcactctccc 80 468 80 DNA Rat 468ctctcacctt gaagagagga gagcgcaaga agagatcagg agaacgggca tggccagctg 60cccgggcctt caactctccc 80 469 80 DNA Rat 469 ctctcacccg gggagagtagtgaactggtc cttgatgaat ccttcaaatc cagactgggt 60 tgttttcaag acactctccc 80470 80 DNA Rat 470 ctctcaccct cacatgtctg tcttggatat cgtcgtccggtgagggcttg ttgcagtcac 60 gaacctggaa aaactctccc 80 471 80 DNA Rat 471ctctcacctc gaaggtatca tcaaacacaa ctctgcaggt cttcccattg ttcaggatgg 60tcttagcaga gcactctccc 80 472 80 DNA Rat 472 ctctcacctg ggtgggcagatctcctccaa aaatatagat gtcctccccc agcgacattt 60 ccactgagtg ttactctccc 80473 80 DNA Rat 473 ctctcaccca gcctctgcag ggcctcgtga tgggccccgagcttcagaaa cactccaatc 60 accgccagtc caactctccc 80 474 80 DNA Rat 474ctctcaccgt tgaggggtct ataatggcca ggggagaggg ccaagggctc aagaagcctc 60tcctgctgaa acactctccc 80 475 80 DNA Rat 475 ctctcacctg cgtggttgtctccaccttcg gactggatgg gctccacgat gatcccagcc 60 actgttctct tcactctccc 80476 80 DNA Rat 476 ctctcaccgt gaaggggttt tctcttggca ggcgcagtgtgcttggtggt ggcttttctc 60 aggatcttgt tgactctccc 80 477 80 DNA Rat 477ctctcaccga aaaataaggt gctttgggaa tctgcgcagt ctgtgtcctg ccggcccaag 60agagaaaccc acactctccc 80 478 80 DNA Rat 478 ctctcaccag acctcaggcttcagaagctg cacctgctgt agtcccgtat gtaggcctga 60 ggtgttgatg gcactctccc 80479 80 DNA Rat 479 ctctcaccgt aggtgcaacg acattcttag gcttcaggctcatagtcttg gtagccttcc 60 tctgaaggca ttactctccc 80 480 80 DNA Rat 480ctctcacctc agcttgggta ctcagccggt acctctggcg tttccagctg cgcttgggac 60tgaagatatc tcactctccc 80 481 80 DNA Rat 481 ctctcaccca tttggatgaaacattttgga taaaaaccta acggttggtg gtttatttgg 60 atattcttca gaactctccc 80482 80 DNA Rat 482 ctctcaccca tggcaaactt cttcccactc tcatctaggatggctgcatt gacgtcctct 60 ccactctcac tcactctccc 80 483 80 DNA Rat 483ctctcacctc ccttaccact gcgcttaatt aagatggccc tcaccctctt gagaaggctg 60agaatgcttc agactctccc 80 484 80 DNA Rat 484 ctctcaccct ttgtgaggctggtgcttgcc acatttctta cagaaagtcc ttcgggtttt 60 aggtacgttg acactctccc 80485 80 DNA Rat 485 ctctcacccg cagagatggg cttggcgctt gtctaaggctttggcagctt cgcgtatgcc 60 acgtgctagg ccactctccc 80 486 80 DNA Rat 486ctctcacctt atccagcatc cagtgttttg gagccgctac gcgcttcaga tgtttcttgg 60gaccacgagc caactctccc 80 487 80 DNA Rat 487 ctctcacctc aatgtgttcctgaatcccaa agccaaagtt gccggtatct gagaagttat 60 tcttccgtaa ttactctccc 80488 80 DNA Rat 488 ctctcaccga tggtgcgtgc cattttcttg tggataccagccaccctgag ctcctccagg 60 ctgaagcccc tgactctccc 80 489 80 DNA Rat 489ctctcaccct tcaaattcat ctgcattgaa cttggtaaaa ccccatttct ttgagatgtg 60gatcttctgc cgactctccc 80 490 80 DNA Rat 490 ctctcaccag gcagagggcaccacctcaat ctgggcctgt ctgttctgga tggtcagttt 60 cactgtaatc ctactctccc 80491 80 DNA Rat 491 ctctcaccga ggtacaggtg ccgtgctgtg gaagcagctcgtgtgtagaa ccagttctca 60 tcatatgggg caactctccc 80 492 80 DNA Rat 492ctctcaccct tatggggcat tcctttttga acagtgccca ttcccttgat gtctacaata 60tcacccttct tgactctccc 80 493 80 DNA Rat 493 ctctcaccct ctccaacagtgacaatgtcg ccaatctgga cgtccctgaa acagggggac 60 aggtgcacag acactctccc 80494 80 DNA Rat 494 ctctcaccaa gcttcagcat ttccttggtg tcaggatctacctcaatgat ctcctgatct 60 agggctgaga ccactctccc 80 495 80 DNA Rat 495ctctcacccc agaactaccg ccttcaccat ggagctccat gagttttccc aattcaaact 60tgggtttctt caactctccc 80 496 80 DNA Rat 496 ctctcaccta agacctgaccatttcccttt aaagcaacag ctctaagaca aaatcttgcc 60 aacctgaggc cgactctccc 80497 80 DNA Rat 497 ctctcaccag gggtccccag gcaggatctg ccagtggtcaaacacacact gggggaaggc 60 ctggccacca gtactctccc 80 498 80 DNA Rat 498ctctcaccaa gaacacagga gtaaaggatc tactcagagt agctctccgg atcgctaact 60ttaaatcaga caactctccc 80 499 80 DNA Rat 499 ctctcaccac tgtggcgactccgggtctcc atagtcccag cgcctccatg atgactgtta 60 ttggctgcct gtactctccc 80500 80 DNA Rat 500 ctctcacctc ttcatgatct gttcagagct ggtgggcccgccgcctccga gctgctcccc 60 ggacccgtcc atactctccc 80 501 80 DNA Rat 501ctctcaccat ccttccgggg aaagaagctg caggtaccaa tagcacttcg cgtcccggct 60cccgccccac ggactctccc 80 502 80 DNA Rat 502 ctctcaccag cctccaaggagctgatctga ggaaaaacct tgtgagtttt gaggccaagg 60 gaactgagat gtactctccc 80503 80 DNA Rat 503 ctctcacccg cgtgcgtaga acttcatgca ggtgtgcttcaggcagggct tacactcttc 60 ccagagggcc atactctccc 80 504 80 DNA Rat 504ctctcacctc cttgctggga aagggactca acacaagcag agggtgtctt ccagatcatt 60ctgtgtaatt ctactctccc 80 505 80 DNA Rat 505 ctctcaccac ttcgtgcaggacagactgtg tctcttgggc gtctgctgca cctcggccca 60 ccatcggtgg tcactctccc 80506 80 DNA Rat 506 ctctcaccgg ggcccagcca tatgaaccca aattggtgaaaactggagga ggcatccctc 60 aggaagggac atactctccc 80 507 80 DNA Rat 507ctctcaccgc agtcgaagtc actcccttta taatgctggt aatttaagca ttccacagca 60tcagattcga gcactctccc 80 508 80 DNA Rat 508 ctctcacctt gttagctacttcacaggtcg gggaaagcag atcagatagg ttttgctcct 60 ctttgtttcc atactctccc 80509 80 DNA Rat 509 ctctcaccaa gacatacaag gagtcatttc atccatccccaactcccaga agtcatttca 60 accctgtggc ccactctccc 80 510 80 DNA Rat 510ctctcaccga aatttaaaaa ataaaaagaa aatgaaaaag aaggttggat ccatcggtct 60tctgtgctag tcactctccc 80 511 80 DNA Rat 511 ctctcacccc agactgtaccaggcaaggtt agtggctatt gaaaatacca ccaggacagg 60 gctagctaaa gaactctccc 80512 80 DNA Rat 512 ctctcacctc tgccacagag aagacccctg gggagggcaccagaaagagg tctgtgttat 60 acagctccat gtactctccc 80 513 80 DNA Rat 513ctctcaccgg atagtaaagc ctttctcgct gtcccaggaa attctgtttc catccggaac 60aaatctcttt tcactctccc 80 514 80 DNA Rat 514 ctctcacctt gggtaggggcctggaagtgg ttgagggcgg ggcagacatg gcttccagat 60 atttccctga aaactctccc 80515 80 DNA Rat 515 ctctcaccgc tcagaccagg cgctccagaa tccgctgaagctcggctcgg ccatgcgtgc 60 tcgaacagcg aaactctccc 80 516 80 DNA Rat 516ctctcaccgc cacgggccta cgagcccatg caacatcgtt cccaggctca tgtggtctgc 60gcctcaggaa atactctccc 80 517 80 DNA Rat 517 ctctcacctg caccagggctccctgctctg tgggagggtg agaagccagt catactttca 60 gggctcaatg gaactctccc 80518 80 DNA Rat 518 ctctcaccga ggggcctggg tagctgggag tgaggggcctttgggagaca tgttcacatc 60 tgtgccatta ccactctccc 80 519 80 DNA Rat 519ctctcaccaa tttctcagca gggtcctatt gaaacagtgc tcgtggttgg gtccataaaa 60atgctggaga caactctccc 80 520 80 DNA Rat 520 ctctcaccac tggcagtcagcagtctcgtg gctgaactcg gggaccccgt ccttcagcag 60 agggtcacag tgactctccc 80521 80 DNA Rat 521 ctctcaccat ccacttcaca ggcaggaaag tgctgccctttgacacgtag ttagaatcgt 60 gcatgatgtc tcactctccc 80 522 80 DNA Rat 522ctctcacccc atcatgcgct gagtgatgcc ctcgattctg tttcctatgt tgctgctgga 60atccaggatg aaactctccc 80 523 80 DNA Rat 523 ctctcaccgg gaagtgcgcactgcctcacc ccggagagcc gctccttgtc ttgttaacac 60 tgtcagagcg aaactctccc 80524 80 DNA Rat 524 ctctcaccaa cagaagcagc tggagaagac gatcatggcgacaatacaga cagccataga 60 gatgagcaca gcactctccc 80 525 80 DNA Rat 525ctctcaccta tttatggaat catctagaac ttcaggggcc atataccttt tagtgcctac 60tctgtggttt ggactctccc 80 526 80 DNA Rat 526 ctctcaccct tccagaccagggtgttgttc tcccccatga tgcccgaagg gcaggtcttg 60 acacagtggg gaactctccc 80527 80 DNA Rat 527 ctctcaccag acagagtagg gtccaggcgc aaggacagcccgaagtcaca caggcaacag 60 gtcaaatcgt tcactctccc 80 528 80 DNA Rat 528ctctcaccgt caatttgatt taaaaaaaaa ataaataaag gcttccccgc tgggctttcc 60actgggaagc ggactctccc 80 529 80 DNA Rat 529 ctctcaccca tccaaagccttgtagaagcc tggccgacaa gcttggcata tgaaccctcg 60 ttcttcatac ccactctccc 80530 80 DNA Rat 530 ctctcaccaa agggtccgga gcgccgcaca caggagaaggcacgtccaga ggggcccctt 60 tagaggtgcg gaactctccc 80 531 80 DNA Rat 531ctctcacctc aatgtaggtt ttggtgcctg gaaatttaaa atgaaagtag agttcttcat 60ccccttcttg gtactctccc 80 532 80 DNA Rat 532 ctctcaccgg accagggcctccagaacgct cacgacgtga gagaagcgag gccgctgggc 60 acggtccttg tgactctccc 80533 80 DNA Rat 533 ctctcacccg tttctagctg gctctcctcc ttggaactattccccaaggg tgtgggtgac 60 agttttcgcg gaactctccc 80 534 80 DNA Rat 534ctctcaccgt agcaccagca gagacatgac agacatacag gccaccacca ccggagtgaa 60gagggactca tcactctccc 80 535 80 DNA Rat 535 ctctcaccac gtcttggtaaaactccatga acttggtcgt ggcgttgagc atgcaggtcc 60 tgtgctgctc gcactctccc 80536 80 DNA Rat 536 ctctcacctc ttgagggcct tgcgcttctg gtggcccttggagtgtgaca gcttggagat 60 gatgatgcag taactctccc 80 537 80 DNA Rat 537ctctcaccac gtaaacagga taggcagtgc caagcacgag aggaggtcgg ccaccgccag 60attcagaaac caactctccc 80 538 80 DNA Rat 538 ctctcaccta tcaggtgtggaaagttagaa cggttaaaaa acaatccctc tcccatgcgt 60 gcacaccttt ccactctccc 80539 80 DNA Rat 539 ctctcaccac tcaggcagct gttggagaag gctgccaggttgactatgtg gcctgtcaag 60 gggtaggcat gaactctccc 80 540 80 DNA Rat 540ctctcaccct gagccatggc agggggaaga tctccttagg atctgctggc cttgtccgaa 60aggccatggg ctactctccc 80 541 80 DNA Rat 541 ctctcaccgt ccccgagagattccccaagg atggtgattt ttcatcatag caacagccgc 60 agccagggca tgactctccc 80542 80 DNA Rat 542 ctctcacctc cttccatctg aagcttttcg gggaagaaaatcagagcagg agtccctgta 60 ccaaagcact atactctccc 80 543 80 DNA Rat 543ctctcaccac gaagatccga tttctctttt gagtaggaat tctttcattt gtatttgaat 60aataccattc caactctccc 80 544 80 DNA Rat 544 ctctcacctc tacgatgctgggagtggagt ggctcccaag tctctgtgac tcaaggcttg 60 ggagctgtca ggactctccc 80545 80 DNA Rat 545 ctctcacccc cagaaggtag ccagcgctgt tcagggtccagcctctcttc tcctttgttg 60 gcatcccgag ccactctccc 80 546 80 DNA Rat 546ctctcaccgt tgcgaggacc agcagcgcca tgaggctgga gtagagcaca gagaagccta 60ttaccgagaa tgactctccc 80 547 80 DNA Rat 547 ctctcaccag gcgttcgtgcccagtcattt cttttgccat taaagatagg aaaggctact 60 gcatgatttt gtactctccc 80548 80 DNA Rat 548 ctctcaccgg ttcataagaa ggaagaggac aatatgaggtcacccactcc aagcagcggg 60 tgcttcatga gcactctccc 80 549 80 DNA Rat 549ctctcaccgg aagcacgctg tctaccagca gagatgtatc cgagacgatt tcagcagcct 60ctcttacgat tgactctccc 80 550 80 DNA Rat 550 ctctcacccc accctgactcctgggaagtg cagtgtgggt aaggggtgcg taaccaaggc 60 agccctaaga ggactctccc 80551 80 DNA Rat 551 ctctcacctc agttccgaag ggccagttga aacggttcccgatgttccct tgccagaagg 60 gcaggcccag gaactctccc 80 552 80 DNA Rat 552ctctcaccgt gtccggggca ggacatgtgc caggtctata tccaggggct cttaagtgcc 60tatatctacc tgactctccc 80 553 80 DNA Rat 553 ctctcacccg ggcggcagcagcttcctgcc ggtgccattg ccctccacgc tacgcagcgc 60 gtactcgatg gcactctccc 80554 80 DNA Rat 554 ctctcaccca tggtgccttc gccatcgctg gaggcacgcaccctgaagga acctggtttc 60 ccacccacca taactctccc 80 555 80 DNA Rat 555ctctcacctg tacttatttt aacaagacaa aactacagta aaaatatatt tttaaaatct 60ccaaacttca ctactctccc 80 556 80 DNA Rat 556 ctctcaccca gcaaatgtaattctggcatg ttctctggga gcacttcaga cctgagtggg 60 ctcccacaag tgactctccc 80557 80 DNA Rat 557 ctctcaccgc ttggaggatc atgcagcgag tgctttatgggcaaaaatca gagtctatga 60 atagtatcat taactctccc 80 558 80 DNA Rat 558ctctcaccaa tttgtgttct gacacagcca ttagctcctg ggaagtacca ttatagttca 60ggacaaccct gaactctccc 80 559 80 DNA Rat 559 ctctcaccgc tgcaatggcttcaaccctag taatggggaa ggtggaggtg gtccagggtc 60 ctgagtagga acactctccc 80560 80 DNA Rat 560 ctctcaccac gtctctgggg ttcaggcgct cttgattgcgccttttaaac ttcctgagag 60 caacgccaag gaactctccc 80 561 80 DNA Rat 561ctctcaccgt taacaacccc cccagcccca aagagcccca agttaccaca tccagcttca 60actccaacac tgactctccc 80 562 80 DNA Rat 562 ctctcaccgg gtgagggaggcggacaatct gagaagaggg gaccctgagt agggtccgac 60 accggcgggc ccactctccc 80563 80 DNA Rat 563 ctctcaccat gcattttgaa acaattccac tttctgtggaaagccagagc ataacatttg 60 aaggtgtaca tgactctccc 80 564 80 DNA Rat 564ctctcaccta gcgagggtct ggatggtgtg agacccagtt agggcaatgg agcagtttct 60gcctccagat ttactctccc 80 565 80 DNA Rat 565 ctctcaccag gtgagatccaaggatgaggg tgtggccttg gggccttcct cgggctcctc 60 ttcatgggct ccactctccc 80566 80 DNA Rat 566 ctctcaccca ataatgaagt caaacgggaa cagccacaatgtcctaagtc acttctgcac 60 taggctatgg gcactctccc 80 567 80 DNA Rat 567ctctcaccgg agctcggtgt ggccactgca tcacttcact tgggaggggc gtttaagcag 60agggccgggt agactctccc 80 568 80 DNA Rat 568 ctctcacctc ctccatcactggattgcctg tgacaggtgt ctgtgctcag agttctttgt 60 tgacacctgt atactctccc 80569 80 DNA Rat 569 ctctcaccaa ctgcaggaat gacatagatg attcccgggtgaaaccagtt gtccatcttg 60 gggggactca gaactctccc 80 570 80 DNA Rat 570ctctcaccct ccagctcatc gtagctggag acttggatga aaggacacca gcgaaatgct 60ctcttgaagc ctactctccc 80 571 80 DNA Rat 571 ctctcacccc aaaacttgccggatggcagg tcctatggta ggtccattga cttcctccag 60 gtggcagtaa ggactctccc 80572 80 DNA Rat 572 ctctcaccgg ctgctggcca tcctttcctc ctgaagagctttgccttctt gccatcgcct 60 ccaacgccgc agactctccc 80 573 80 DNA Rat 573ctctcaccgt cgtaaacaaa tctatccctc actgctaaat agcacccaag ggcaagaggt 60aatctgcttt atactctccc 80 574 80 DNA Rat 574 ctctcaccca ccacgttggtagagtctgtg gccaggaagt aggatgcagt agccgcgatg 60 gatatccaaa tgactctccc 80575 80 DNA Rat 575 ctctcaccta gactggctgg gctgccggag gcttcgaagagggtgacaga cagccaggta 60 ccggtccagt gtactctccc 80 576 80 DNA Rat 576ctctcaccgc caggagctga atgaccatct ccaggtggtg agacctgcct tgcctgtgct 60gctgacttct aaactctccc 80 577 80 DNA Rat 577 ctctcaccct cggcagggcctgcaatttgg tggaagaaat atccaggatg ctgggccccg 60 tggccccctg gaactctccc 80578 80 DNA Rat 578 ctctcacctg tccagttcga tgcttccgaa gtgttagcatctctggagtt gaagtcatta 60 tggtagagcc tcactctccc 80 579 80 DNA Rat 579ctctcaccct ccaccatcaa tgctggaacc tcaccagggc tcttcttgga tatcctgttg 60ttgtcgtgca tcactctccc 80 580 80 DNA Rat 580 ctctcaccgt agtcatttaaagtatctata cattattatg taaggaggaa gtaaagaatc 60 ccaaatggga tgactctccc 80581 80 DNA Rat 581 ctctcaccgg gagggtgaaa tggttttcag aaagtagtggtcttggtgaa agggactcgc 60 ttgcaggcag gtactctccc 80 582 80 DNA Rat 582ctctcaccgg gcatggaaga gaaaaacttc cattcacaga ctccatctgt cttggagatc 60ttatagaaag tcactctccc 80 583 80 DNA Rat 583 ctctcaccct agggctccaggcgcgctctg gggccagcag gtcccgatct ggtccaaggt 60 cgtgttgcag taactctccc 80584 80 DNA Rat misc_feature 67 n = A,T,C or G 584 ctctcacccc agtttcctgtactgctttct catgtcatgc ccaacagcat tggtctccca 60 aattttngta gcactctccc 80585 80 DNA Rat 585 ctctcacctt ggtgagcaca aacctcagtt caatggcgttccgggggagg tcggtcggaa 60 tctctgtcac ctactctccc 80 586 80 DNA Rat 586ctctcaccag gatcaattat ggagttcact gataaaaatc taagagctcg gaggtcccac 60ttttcctttc ggactctccc 80 587 80 DNA Rat 587 ctctcaccgt gagggcattagaagcttgtc actgcttatc agtttctcat ttaatatgga 60 tgcttaggca taactctccc 80588 80 DNA Rat 588 ctctcaccgc tgtagtcaca caatgcctgg tctccgggccactggccctt catgtatgtg 60 gcgatggtca ctactctccc 80 589 80 DNA Rat 589ctctcacctc tcatccttca gagtggaaag ctttctctgg gagaaggatt caaatcctag 60cagtttgaac acactctccc 80 590 80 DNA Rat 590 ctctcacctg gggagcggctcaggggccca ggcgggctct cttcctctag ggatgcagag 60 cccgtgtcca gcactctccc 80591 80 DNA Rat 591 ctctcaccga tgggtcgagt tgctcccagc gtcgctgcgacaggtacacg aggatgacca 60 ccggactggt caactctccc 80 592 80 DNA Rat 592ctctcaccaa tgatgaatgc caacacaaca gcagctgcca tcttcaagac ttggtcacgg 60gtaattctgt tcactctccc 80 593 80 DNA Rat 593 ctctcaccgg tgactgggacgttgggctga tgcaggctct ggcgcacctg ggcacagtgc 60 tgtaacctca caactctccc 80594 80 DNA Rat 594 ctctcaccga gccgccgcgg gtccacggcc ggtaggtgtaagcgcacgcg cctagacggc 60 gacggcggcg gcactctccc 80 595 80 DNA Rat 595ctctcaccat atcagtggcc aagagggctg cgtagataaa aagagggcac agtgaggtag 60tgtcccaact ttactctccc 80 596 80 DNA Rat 596 ctctcacctg aatgcatctcagcaaactct cagatcccag aagacaacac cgtggaggcc 60 tggaggaccc ccactctccc 80597 80 DNA Rat 597 ctctcacctc atgtttgatg atacactgat catcacggctatttccttgg tgcctccaga 60 agagaggagg caactctccc 80 598 80 DNA Rat 598ctctcacccc cttggcagat ggatcttgtt cctctggaca gctgggcaga ctacaggggt 60ttgtgaggca ggactctccc 80 599 80 DNA Rat 599 ctctcacctg tccagcccttgtctgactcc attatgttgg ctgctgtgtg cagaacttcc 60 ttgagcagct tgactctccc 80600 80 DNA Rat 600 ctctcacctc accatgattg cttcaggctt aagcaaccagagtttcaaac tgagaattag 60 gaagttaaca gaactctccc 80 601 80 DNA Rat 601ctctcacctc cctacagtct gtcccagaga agttttccaa gttgtcccac tgctgtcccg 60ggactagagg gaactctccc 80 602 80 DNA Rat 602 ctctcaccca ttctcccccgccgccttgcc caacccattc tctgtggtcg gggacgcgcc 60 gtcggggccg gcactctccc 80603 80 DNA Rat 603 ctctcaccgc tgccaccgct gccgctagac agctgctgaagctccagctt cctcgggggc 60 cgggacttct gcactctccc 80 604 80 DNA Rat 604ctctcaccgc acgatcatga tggccgccct tttgggagtt cttttagcag aatagtccac 60cgcatcagtg atactctccc 80 605 80 DNA Rat 605 ctctcacccg cctagaaccaagcagcccag agatcttcct ctcacctcag tggactctgg 60 ggccaagggt ctactctccc 80606 80 DNA Rat 606 ctctcaccat ggagcaggaa agagggactc catgctgtcaccctgcaaag ccactcagta 60 cagttcctcc aaactctccc 80 607 80 DNA Rat 607ctctcaccca gggcaaaaag tacaaaaact acaaacatgg tgacaaagtt cctgaagtcc 60tgcggcttca gtactctccc 80 608 80 DNA Rat 608 ctctcaccga agacacagaatgtagagtta gagttgtggt tgaatttcct tttctctatc 60 acatcaacta tgactctccc 80609 80 DNA Rat 609 ctctcaccaa ccgatgttag caggagatat atgtgaattacgtgtgagtt agatgtgtgg 60 tggtgtaagg taactctccc 80 610 80 DNA Rat 610ctctcaccag atgatcaaca ccaccaggat cagaaccagg gttttggcca gcctaatgtc 60catgcgggct tgactctccc 80 611 80 DNA Rat 611 ctctcaccac cttagagtggttgtagtgga agaggtcgtt caggatgcta ttggccagga 60 agggcaagga gaactctccc 80612 80 DNA Rat 612 ctctcaccag ggccagcagc tcatgttgtc ccaaagcccctcacagcctg atgctgtctc 60 cgggcccagt gcactctccc 80 613 80 DNA Rat 613ctctcacccc agcagtagca gcagggctga gagcgccagg ctgagcacgt ggtagcgcag 60gcgcccgggg cgactctccc 80 614 80 DNA Rat 614 ctctcaccgg cagccagcagaccgcgaagg cgaccacgat ggagagcagc atgacgttga 60 ttcgtttggt ctactctccc 80615 80 DNA Rat 615 ctctcaccct agcgtgcgca atccggtgcg tgcagagtggttttaaatgg gatattcaaa 60 catgtcccta ctactctccc 80 616 80 DNA Rat 616ctctcaccag gctgtgtgtg gtgtgcctct gagcacagca aagcaggcta cgcaactccg 60aggagacact gcactctccc 80 617 80 DNA Rat 617 ctctcaccat gctaaggctaggcacagtac cttgacagtt cacaaagcct tccatgccag 60 gggcttcctc acactctccc 80618 80 DNA Rat 618 ctctcaccct tgtggaggtc ctgtgtgagg ctaagaaaggtaaagtgctc tgatggataa 60 ggggtagagg agactctccc 80 619 80 DNA Rat 619ctctcacccg gtcggaggca gcatctgtct ccttccagat gggtttaggg gagagagggt 60cacaacggaa ccactctccc 80 620 80 DNA Rat 620 ctctcaccat ggcggatgaacgggcagatg gaggaatggc ggcactccag atgcacagag 60 aaaagcctct tgactctccc 80621 80 DNA Rat 621 ctctcacccc aaaggtctcc ttaagttcca caagactgtgaaacactggg aggggagaac 60 agatggcaaa gcactctccc 80 622 80 DNA Rat 622ctctcaccgt aaggtcaata aacataacac gttgagactt tttccccacg tcaaaagtca 60taaacttaca ttactctccc 80 623 80 DNA Rat 623 ctctcaccag gaccatctctgggaggggaa gaatgaagtc ctgacccagc tgggacagtg 60 gctactcccc atactctccc 80624 80 DNA Rat 624 ctctcaccag tagtctggtt gcaggaatca cacagagctaaagttacgtt tgtaatgaaa 60 aaggggcacc acactctccc 80 625 80 DNA Rat 625ctctcaccag ggatcaggta aaccagatta cagtccttgg agatggagcc tcggggctca 60tggtggctgg aaactctccc 80 626 80 DNA Rat 626 ctctcaccgc agcgaccactttgtgatcag ccgatctctt tcccaggacg aggacccagc 60 tatagttagc atactctccc 80627 80 DNA Rat 627 ctctcaccgg cagggctctc ctgtgtgtgt gtaggggtgagtaagggcac ctgcccctat 60 tcctcattcc tcactctccc 80 628 80 DNA Rat 628ctctcaccgg agggggcaga ccaggagacc ccatggaaga actgatgact gggaaggggg 60agcccagtgg cgactctccc 80 629 80 DNA Rat 629 ctctcaccag ttcctgaagctggaacggct cctcccctca cgcgccaaaa gccacgccca 60 acctgaggtg gcactctccc 80630 80 DNA Rat 630 ctctcacctt ttgtgtatgt gtttgtgtgt ttttaaataacaaaaggata gtaatcccca 60 tattgactag acactctccc 80 631 80 DNA Rat 631ctctcaccct aacactgaaa ccggcacata agaaacacat tcaatctcag cttaccgttg 60gggtgaccaa ccactctccc 80 632 80 DNA Rat 632 ctctcaccac ttaagcaggacaatctggtc atcggaggtg agatccctga atcctgggat 60 catcttggca aaactctccc 80633 80 DNA Rat 633 ctctcacctc caaacagagg tgccacgcgg gcatgtcgagaaccagcgac agcaaccagc 60 aaccagaaca agactctccc 80 634 80 DNA Rat 634ctctcaccac tacaggtata aggctgattc agagtccagg cctgttccag atatatgctg 60ctaccgggca caactctccc 80 635 80 DNA Rat 635 ctctcaccat gtagtatctgcagctagctg cccacctgcg ggaagctcca gggaagaaaa 60 cttgcaggga ggactctccc 80636 80 DNA Rat 636 ctctcacctc aatgaagccc gggatctttt ccgcccacttccggatgacg tccagggagc 60 cagagagcaa gtactctccc 80 637 80 DNA Rat 637ctctcaccaa gtgacattta ttgtacaaat ggttaataaa aagacacatt ataaatatat 60atgtaacctg ctactctccc 80 638 80 DNA Rat 638 ctctcaccgc ccacgcaaccccgcggtggg tcagtgatat tctccttctc taggagttcg 60 tgaactgcct gtactctccc 80639 80 DNA Rat 639 ctctcacctc catctacagg aagtgcaggc tggtagagcaaaattgaggt aaagcccagg 60 gagcggacgc tcactctccc 80 640 80 DNA Rat 640ctctcaccag gtaggaatgg ctttaaactg tcattctcat ttttacagca acttctgtgg 60tgcctgccaa gaactctccc 80 641 80 DNA Rat 641 ctctcacctc ctccaggcgcacgccggaac gaccacctcc tccagacgtc ttgttcaagg 60 ccaaagccaa ggactctccc 80642 80 DNA Rat 642 ctctcaccct gcatgattct ggctgggatc gtgtctgtgtagttcaccag ctggaaacct 60 gtcacattgc gtactctccc 80 643 80 DNA Rat 643ctctcaccga aggcctggac ttgaggggag ggagggaggt gtaagagcta tctctggtgt 60tccgaggcct ctactctccc 80 644 80 DNA Rat 644 ctctcaccga ggagctgcaaatgaatgaga cctcacttgg agctgggagc tgagtggtca 60 gggtggagga agactctccc 80645 80 DNA Rat 645 ctctcaccca gtacttcctg taattgacag ccttgccttgctcctcatga catcactcaa 60 ggtcatcccc taactctccc 80 646 80 DNA Rat 646ctctcaccgg cacaaacctt tcatattcgt tccagtaacc agcttttcga gaacccgaga 60ctttcatttc atactctccc 80 647 80 DNA Rat 647 ctctcaccag attgccagtctaaaagcagt gtattcctgg tctgtgtttc ggatgaagag 60 accacctatt tgactctccc 80648 80 DNA Rat 648 ctctcaccgc atcttctcgt tgttggcatt agcagctttctcagctgctt tcttttggcg 60 ctggggtcct ctactctccc 80 649 80 DNA Rat 649ctctcaccag gccggccaca gaagccagtc cagcaggaac caaagctgcc agtaacctca 60cacttctgat tcactctccc 80 650 80 DNA Rat 650 ctctcaccgt tcgatatgaaatatggaaat aaggaaacct tcgaattctt ttaagacaga 60 ggcagtaagg caactctccc 80651 80 DNA Rat 651 ctctcacctg ttgttgttga agtaaacatg gtgtgataataaaaactgac acacaagatc 60 gaaaaccaac tgactctccc 80 652 80 DNA Rat 652ctctcacctc catgtttatg ggatcaacag gtccaatgct gtttacataa acatcagttt 60caattactgt ggactctccc 80 653 80 DNA Rat 653 ctctcacccc attatatacatactatacat attgggtcat aaattagcag ttattagtca 60 gactgtacaa gcactctccc 80654 80 DNA Rat 654 ctctcaccat attgctgaat tcctggtgtc accaacgctgcctgcaacct cattcatttc 60 attgtgaaca tcactctccc 80 655 80 DNA Rat 655ctctcaccac gtcaatgtgg atgcggcctc ttctccagga gccagacttg cactcctcat 60agcagcagaa gaactctccc 80 656 80 DNA Rat 656 ctctcaccac gccgaccgcaaaggagctgg ctgcagtgtc ccgtgaggag agggcgatgc 60 ggcagacgaa agactctccc 80657 80 DNA Rat 657 ctctcaccat ttagaagaaa aaccgtcctt tcctccaacttgatcttgaa gctttctgaa 60 atatttgcca caactctccc 80 658 80 DNA Rat 658ctctcacctg gaaaggtaaa ctatccagta cacaaggttg aatcctgcaa atgctactgg 60gaagagaatt cgactctccc 80 659 80 DNA Rat 659 ctctcacctt gagcgaggagattcagacac acaaaggcca aatagagact gtagctcata 60 ttgaaggagc tgactctccc 80660 80 DNA Rat 660 ctctcaccac aggggaagat gccctgtgac ttaaccatggatcctgtctc cagtctccgg 60 tcagcgcttc tcactctccc 80 661 80 DNA Rat 661ctctcacctg aaagctaatt cttcaacatt aacaggttca ttttccacag tttcaaaaat 60ccctccgatc ctactctccc 80 662 80 DNA Rat 662 ctctcaccgc cgtcataggtccaagtgccc agcttcatgc tgcagttctg ctcatcgaag 60 ggaaagtggg tgactctccc 80663 80 DNA Rat 663 ctctcaccgg ctgcagcacc caggttctgg cagagggcggctgcggtcca agttccatgc 60 cgatgcctct gtactctccc 80 664 80 DNA Rat 664ctctcaccgg gtggaggaca ggagccaggc gatggacagg gactggcctt ctcaacctct 60gatgtcttca agactctccc 80 665 80 DNA Rat 665 ctctcaccgc accacgtccgggagccaaac agattcagca gtgacacgga gagactcgat 60 gccatcgtgt tcactctccc 80666 80 DNA Rat 666 ctctcacctg ccgatgctgt gtggccggtg gtgatggtgatagtggtctt ggtacacatt 60 gctgtccttc agactctccc 80 667 80 DNA Rat 667ctctcaccta cacatggctg ctggcacatg tcaccacatt aacaactcta ggaacacaac 60acacagctag ttactctccc 80 668 80 DNA Rat 668 ctctcaccca gttattcaatggatcagtaa acaggtagct tctctcccgt gaggtcgata 60 aaaacccatt ttactctccc 80669 80 DNA Rat 669 ctctcacctt ccaaacatgc cgatgtactt cccttttttctgattctttt gagaaaaatt 60 tctgtggtat gtactctccc 80 670 80 DNA Rat 670ctctcaccca gaacatttac aaaaatgtct tccagggccc cttgggtgtt atctctccca 60gggacccaca ggactctccc 80 671 80 DNA Rat 671 ctctcaccat caaagggtaaaatttgtctt tatcggataa cagccattga tctgaccaca 60 gagctcagaa ctactctccc 80672 80 DNA Rat 672 ctctcacctg agcaggtggg ggttgtctgc tttgctgggcttgatgctga caggctggaa 60 gcccgaggtg agactctccc 80 673 80 DNA Rat 673ctctcaccaa cttgccctgg ggaatggctc catttggtac caacaggctg acccctgtgc 60cgggaatggt caactctccc 80 674 80 DNA Rat 674 ctctcacctg aaaatcacagtacagtttga atttttttca gttaatcaag gatatttaaa 60 acgatattca ctactctccc 80675 80 DNA Rat 675 ctctcaccat agatgagaga ccccttcccc agcctcgagccccacacccc acctctgctc 60 cttctgaact ctactctccc 80 676 80 DNA Rat 676ctctcaccct cagtccgagc ccactatgtg tggctctctc ggggtagatc ctgatgaatc 60ttgtggagag agactctccc 80 677 80 DNA Rat 677 ctctcaccat tgtttaaagctcctcccgga gggtcttcac ttctacatga cagccatcca 60 ccgggaatcc acactctccc 80678 80 DNA Rat 678 ctctcaccgg gcggagggcg caagtagcac cccgggggcgcccgtggccc catctgtcct 60 gaggttaggt cgactctccc 80 679 80 DNA Rat 679ctctcacctg ccagggccac tgagaaatgc cagtcactgc ttcgaaggac agcagtctgg 60agaacttgca tcactctccc 80 680 80 DNA Rat 680 ctctcacctg ttgcacagccctgtacagag ctctggacac agccaccctg atcccagtta 60 tgttttgcag tgactctccc 80681 80 DNA Rat 681 ctctcacctt caaagtcgtt gtaatcggct catacgtaattttgaaaggg ttcgctaaca 60 taaacccaga ttactctccc 80 682 80 DNA Rat 682ctctcaccag ctaatgacaa cttctatgtg cgatggtttg cagcgcatac gtcacagctg 60accgcgtgcc agactctccc 80 683 80 DNA Rat 683 ctctcaccat cttgcttctctttttttctc ccccgtaact tctaatggcg ctctttttgg 60 tgatagcttt gcactctccc 80684 80 DNA Rat 684 ctctcacctg catgttatgc agtgcataag ccatggaatacactgcatca ataacaaatt 60 gaacctttcc ttactctccc 80 685 80 DNA Rat 685ctctcacctg gcacccacac tcaaccttgc tacttggcgt cctctgcgtt ctcgttctgg 60ggtggggctg ccactctccc 80 686 80 DNA Rat 686 ctctcaccgg tgggtaagtggctctggggg caggcatggg gtgggtgtta ttgcagtgct 60 cccagccaac ttactctccc 80687 80 DNA Rat 687 ctctcaccct gcaccaggcc cccggttctc tgtgctcttgggaaagggtt tgatgaccgc 60 cgtttggttg ggactctccc 80 688 80 DNA Rat 688ctctcaccca gggagacaca gaacatgatt aaccaaaaca tcagaatcag aagggcacac 60agctgaaggt tcactctccc 80 689 80 DNA Rat 689 ctctcaccgg gggtgtcccagggcgtcagc cagtctttgg cacctggcac tgtggctccg 60 ctggccggcg caactctccc 80690 80 DNA Rat 690 ctctcaccag tggtcagagc tccaggaggt ctcagatccttttcccaaag tactgaaggc 60 cagatttctg tgactctccc 80 691 80 DNA Rat 691ctctcacccc atgtcctggg gaaggggctg ttaattaggg cactaaaatc agcctagaaa 60gttgtagcac atactctccc 80 692 80 DNA Rat 692 ctctcaccag agtagaagagaatgggggag atggccacca ccacaatgag ccataccagc 60 acactgacat aaactctccc 80693 80 DNA Rat 693 ctctcaccac acagcggaac gggtcagggt ccggcgggatgcgcatatct gggcgatgac 60 acagacgttc agactctccc 80 694 80 DNA Rat 694ctctcaccga gccgttgata tcatatcaga ggatatcagc ccctttgaac aagctttggt 60ccctctacag ctactctccc 80 695 80 DNA Rat 695 ctctcacccc cagcatgttggagcagggtg catctgtgac ccggcaacca aagagctcca 60 gtcgaagggc taactctccc 80696 80 DNA Rat 696 ctctcaccgg tctcgttggt ctccagagga aagatattttcagaagggat gtgggaccat 60 ttcttctgtc gaactctccc 80 697 80 DNA Rat 697ctctcaccga gattgttcac tagagtggcc agctctgacc cagtgaattt attggcacca 60tacactgctt caactctccc 80 698 80 DNA Rat 698 ctctcacctt gcagctgcatgctttgggat gaaaccacac agatgaccac cagcaggaga 60 atgttgaagg ccactctccc 80699 80 DNA Rat 699 ctctcaccct gaagttacaa ctaatttcta cttctgtctgaggaggtcac cacattagaa 60 ttacaatgct taactctccc 80 700 80 DNA Rat 700ctctcacctc tcacctggcc ctacgagagg gcagaggatg agcaagaaga tggggccatt 60aaaatgttag ctactctccc 80 701 80 DNA Rat 701 ctctcaccag gaaaagtcaggagagcaagt ttcatttccc atcaccacat ggaagggtct 60 tcaagccttg ttactctccc 80702 80 DNA Rat 702 ctctcaccga gcgcaggcac tgggaccgcc acaggaaccctgtctttatg atgcagtcca 60 gggtctgaga gtactctccc 80 703 80 DNA Rat 703ctctcaccga ataatgaaat aagaacctct tagcattatc tttctcagtt tgctcctgag 60gcttctggaa aaactctccc 80 704 80 DNA Rat 704 ctctcaccgg ccgggaaaccatccatctac agcgagtgat gaaaggctga gtgtaccttg 60 acccagcgcc ctactctccc 80705 80 DNA Rat 705 ctctcacccc tcatatctga agtgccttcc agtctggccttcacctgacc actctgccct 60 tcatcttccc taactctccc 80 706 80 DNA Rat 706ctctcaccaa agagggcatt ggcactgccg ttgagttgag ccagttgact cttgatctgg 60ttcatgaggt tgactctccc 80 707 80 DNA Rat 707 ctctcaccac ctgggagtagataaggtaca gcccatctgc tggtaccacc agttggttgt 60 ctttgagatc caactctccc 80708 80 DNA Rat 708 ctctcaccga cgcacccccg atttttgccc ctctggcctctgcgaccttt ccctctggaa 60 ttctctgggc tgactctccc 80 709 80 DNA Rat 709ctctcaccgg ggaggacact ccttccccga actgatgggt ttcgggaagg ggaacgtttt 60cggacattcg agactctccc 80 710 80 DNA Rat 710 ctctcacctg tgggtttcttttcgaaaacg aaactttctc agcttttcca ctaaatcttc 60 agcaacatca caactctccc 80711 80 DNA Rat 711 ctctcacctt ctgaccagtc ttaactactc ttccctcctcaagtgaggca atcaaaaccc 60 atggaggcat gcactctccc 80 712 80 DNA Rat 712ctctcaccgt ttaaatatta aacagggatg tcttgtcccc cagaagggag accagatgct 60gcctgggccc ctactctccc 80 713 80 DNA Rat 713 ctctcacctg aggttgccccgtagaccctg cttgtatagc ttcaggcggg tctgcacaca 60 tgtcggcctc tgactctccc 80714 80 DNA Rat 714 ctctcaccac aggcacagtc ctgccttgaa agccatctgactgcgtctgt ttgccccccc 60 acctccctca tcactctccc 80 715 80 DNA Rat 715ctctcaccct aggatactgt acggacagtt ttccttcttg gtcccgctga ccttgccgtt 60cttttcaatc ttactctccc 80 716 80 DNA Rat 716 ctctcacctg acatactccactttggccac cttgacactg cggtggtgga cccttgaggg 60 ctggcacttg acactctccc 80717 80 DNA Rat 717 ctctcaccgc gagagcagga tgtctagtga gctgcctcagagccttcctg aggaagcttg 60 cccctgagag atactctccc 80 718 80 DNA Rat 718ctctcaccga agcgtgtcaa caagctcccc tccgcacaga gtctcgctgg ggcggtaagc 60agcgatgcag caactctccc 80 719 80 DNA Rat 719 ctctcaccga cagcaggctgcctttgttgc aaagtgcagg gcgaggtttc tgcaacggac 60 tccaggtatc gcactctccc 80720 80 DNA Rat 720 ctctcaccag gatcttgcgg tgacgtggca ttttctgttcctcgggaggc tcctcctaca 60 ttctgtaggt ctactctccc 80 721 80 DNA Rat 721ctctcacctg gtgttatccc acagcatgtc aacaggacag agtttcttgg aatgaggaca 60ttgatcttct tcactctccc 80 722 80 DNA Rat 722 ctctcacctc aagcgaaggtggaaccgttc cagcccacgt tggctgcgtt catgtcgtaa 60 tagttgatgt agactctccc 80723 80 DNA Rat 723 ctctcaccgc agggtgaatg acgagcagga ccggctagggatgcgggcgg agctgcggag 60 ctggaagctc atactctccc 80 724 80 DNA Rat 724ctctcaccgt ccgtaggaca ggccggtaag aaggtaacaa acaaccccgt ttgatgaggg 60ttgggagtgg tgactctccc 80 725 80 DNA Rat 725 ctctcacccc gaacggtaccgcgtcctgca caccgaaata gctcccaact tcacctaagg 60 gagccatcag agactctccc 80726 80 DNA Rat 726 ctctcaccgt gaatcgctga cgcataggta ttatagctgttttcttggaa tctctccctg 60 aacttacagt caactctccc 80 727 80 DNA Rat 727ctctcaccaa tatacagatg agctactcgt aataaaccat ctgcgttgat caatccagtg 60tagccccagc cgactctccc 80 728 80 DNA Rat 728 ctctcaccaa ggaaccgccatctcagaagt agccatcctc tgaaggctcc ttgagctcag 60 atcctcctca gtactctccc 80729 80 DNA Rat 729 ctctcaccag ccagcagccg tccatcttcc ttcatagccaggtaccggtt cgcacacact 60 cccttgatgg acactctccc 80 730 80 DNA Rat 730ctctcacctc acacactgag tactctcctc ggtgactctt atgctctgca tagcgtttcc 60tccgtggtga tgactctccc 80 731 80 DNA Rat 731 ctctcacctc caacccagaacagtaaatat tgcacacatc tacaaataaa tttggatttt 60 tactgagggg ggactctccc 80732 80 DNA Rat 732 ctctcacctt ttttcctcct caacctgagc ggttcccaagcctggacgtc aggcacggcc 60 tgttcggctg agactctccc 80 733 80 DNA Rat 733ctctcacccc agcctccttg tactgatact cggccccagg gagctcgttg ttctgcaatg 60gcaacaagat ccactctccc 80 734 80 DNA Rat 734 ctctcaccag ccccgagggcggcatggggg aggcggcgcc ccacggaact tcggagagcg 60 cgaaccccgg gcactctccc 80735 80 DNA Rat 735 ctctcacctc cttctttctt ctgcttcttc atattccctgaagaatcgtt tccgaagcag 60 gacggcggta atactctccc 80 736 80 DNA Rat 736ctctcaccca ggtaccactg ggtgcgacag aacagtctcc tcacccttat atcccctcct 60tccatgtagt caactctccc 80 737 80 DNA Rat 737 ctctcaccgc taggaatgttgtcgataaga gtcctcacgc acgtagaatt cggatgcaaa 60 caaattgcat tcactctccc 80738 80 DNA Rat 738 ctctcaccga gaaacaagat ggctttctgg ccgtagtgagtccgaggacc gcgcttacaa 60 ctcccgttct tcactctccc 80 739 80 DNA Rat 739ctctcacctc cgtgtctggt ggaggtgagt cacctcaatg gccagcccat agttgggttg 60cttttcccgg gtactctccc 80 740 80 DNA Rat 740 ctctcaccat gtgagaaactcatcagtagg gacagaactt aaattgaaga agaagcgtcg 60 ggaagttttc ccactctccc 80741 80 DNA Rat 741 ctctcacctt gcagcagccc gcacaccgca ttaggggcacacaggacggc ttgaagatat 60 actctatctc atactctccc 80 742 80 DNA Rat 742ctctcaccaa tgtgacaata aaataaatta aaaaattaaa tagtattaaa tcaataggat 60taaattaaat aaactctccc 80 743 80 DNA Rat 743 ctctcaccca ggtgaaacctccaaccccag ctgtgatcat ttcctccttg ggctgtgtcc 60 aattccatcc caactctccc 80744 80 DNA Rat 744 ctctcaccag ctgacctgtt ggaccttgtg gtctttgtggctcaattggt cttaagaaga 60 ctctcattga agactctccc 80 745 80 DNA Rat 745ctctcaccag cgactgactg cgccgatccg gtctatcttc tgcccaaagc agcttgaact 60atgtgccatc ttactctccc 80 746 80 DNA Rat 746 ctctcaccca agccctccagcagctgcctg atcctcacag gtggcacctg tggctgggaa 60 aaggatggcc tgactctccc 80747 80 DNA Rat 747 ctctcaccaa gggtgggtga atggcaatgt ccattgttccttattctttc atttcaccga 60 aggagcagta gcactctccc 80 748 80 DNA Rat 748ctctcacctt ccacgttatt tgagtccttc ttgaggcagt gaaacaggtt ataaaaggca 60aagtatctag tgactctccc 80 749 80 DNA Rat 749 ctctcacctc gagttggcagacctctgcag ctccagcctc atctcgtcct gctcagctgc 60 ctggggcaaa tcactctccc 80750 80 DNA Rat 750 ctctcaccgt gaagatggcg tctgcatgcc gccgcaccctgaagggaggt gaggggggca 60 gtgagcagtg ggactctccc 80 751 80 DNA Rat 751ctctcaccac ctaggagagg cactcaccac ccctccccca acacctggcc acagtgatcc 60tggttgcagg taactctccc 80 752 80 DNA Rat 752 ctctcaccta ttagtcttcttcaactttga tttgtcaaat gtttcaactt ctgataagtc 60 tggtttatca ctactctccc 80753 80 DNA Rat 753 ctctcacctg caggtgctgg gctcggagca cagcattggcaaacagactg gacaagggca 60 tggcaggtaa agactctccc 80 754 80 DNA Rat 754ctctcaccgg tgctcgtgca gaaggcgagc ccacgccgcc cgggacttgg tgtccacacg 60caggtcccga agactctccc 80 755 80 DNA Rat 755 ctctcaccag cctctggtttagcggggtag gcgtccacca gcgctcccag gcaggcgagc 60 aggacaagca gcactctccc 80756 80 DNA Rat 756 ctctcacctc ctatgggaaa tttagctgag ttttcttcatccccaatttc cctcttttcc 60 tgtgtggact caactctccc 80 757 80 DNA Rat 757ctctcaccga caccgcactg tatacgggat ttgctccaat atggcctggg agccaaaagg 60ccaggaagag gaactctccc 80 758 80 DNA Rat 758 ctctcaccct gtagtcagtgttacacttgc cacacttgca gctcagggca acggggtagg 60 agaaataagg agactctccc 80759 80 DNA Rat 759 ctctcaccaa gaagtcctaa gagaaccact gctttctcatctgcagacat tgactttata 60 aaccttcgac acactctccc 80 760 80 DNA Rat 760ctctcaccca gcgtcaatgt cacactcgga tgttgtggat ccttctgggc cagtgcatta 60catcttcttc tgactctccc 80 761 80 DNA Rat 761 ctctcacctg agcttgctgagctaactgct ctgccctggc catttccaag acttccctca 60 gaaggtggaa ggactctccc 80762 80 DNA Rat 762 ctctcaccgc aggcgctgga tgctgctaga cactgggaagatgtgccaag tgctcttccg 60 agcatctacc acactctccc 80 763 80 DNA Rat 763ctctcaccta gagccagcac taagactagc atgcacacac acagtcgagg catcttgtct 60gcagagctgg ggactctccc 80 764 80 DNA Rat 764 ctctcaccaa gcaagatggaaactggaaag gagaggaggg acctagttgc caaaataggg 60 gtggtggttt gtactctccc 80765 80 DNA Rat 765 ctctcaccga atgcatcctt ttttgcttta ctgttgctgaagaagttagt gatcaggtgc 60 gattcgatga caactctccc 80 766 80 DNA Rat 766ctctcaccca atattttcag gtggattcat ttcctcaaag gaaatgattt tgtttttaca 60ggagagggta gaactctccc 80 767 80 DNA Rat 767 ctctcaccga aacagtgtacgtctgtctat agcatggtgt ttcagaggac actgcatcta 60 actgaccacc taactctccc 80768 80 DNA Rat 768 ctctcaccgc aatccttaat cttttggggt ctgtcagcctcaaagaacag gtcattctcc 60 tcactgtcga aaactctccc 80 769 80 DNA Rat 769ctctcaccct ttagttttac aacagttact ctgatattgc tgatgaaatt tccagcgtct 60tccaagtgaa agactctccc 80 770 80 DNA Rat 770 ctctcaccga agtctcttgcggagagaaac ttcatagctg ttcctgaagg gcagatggag 60 ttgactttcg ttactctccc 80771 80 DNA Rat 771 ctctcaccag acgatgtgac cggcagcagc acaaggatcaggggaggaat tccaaagata 60 tatctaaaag aaactctccc 80 772 80 DNA Rat 772ctctcaccta aaacttgatc atttctgaca aggcttggca acccaagtaa cccttaaagt 60cctgcagtaa ggactctccc 80 773 80 DNA Rat 773 ctctcaccat agcggaaaagttgcttggag taattgagca gttttgatat aaactgggct 60 acttcgattt tgactctccc 80774 80 DNA Rat 774 ctctcacctt gttagaagac agagtgctgt ttgcaaggtagagcacgttt cttactgttt 60 cattaagagt caactctccc 80 775 80 DNA Rat 775ctctcaccct ggctgtggtt tactgcaccc tttgggcgtg aaccccatca ttctcctgct 60gctcgttgta gaactctccc 80 776 80 DNA Rat 776 ctctcaccct gcagcctgctcagagccacc acctctgtgg agtagagcga ggcttccagg 60 acgccatcca ggactctccc 80777 80 DNA Rat 777 ctctcacctt gcagtttggc agataaaatt tgtagatctcatctcctgct ttctgttgag 60 cggcacgtaa tcactctccc 80 778 80 DNA Rat 778ctctcaccgg gccaagtcat caggaagctg cctgacaaag gaaggtcctt gtccttgtct 60ttatgggctc tcactctccc 80 779 80 DNA Rat 779 ctctcaccac actggcatctgttcccttgg tctgtggtct ttgtgggaag taagtctttc 60 aaggatcgct taactctccc 80780 80 DNA Rat 780 ctctcacctc acatagagct caaatgacag ccccatgctcctgccagggt cctggaggct 60 cccaccaggt ttactctccc 80 781 80 DNA Rat 781ctctcacccc ggcacggcac aggcggccag cagcaggagg accacgctga tcaccatttt 60ttccgagtct ggactctccc 80 782 80 DNA Rat 782 ctctcaccag cttcttcaggacttcctgca gcgcttcaat ctgcaacaca gcgccgggag 60 cccgaagttg ccactctccc 80783 80 DNA Rat 783 ctctcacctt ttccactgga gaccatcctc accacagcattgtcgcagct aattttcatc 60 tgggcagggc caactctccc 80 784 80 DNA Rat 784ctctcaccgg atggagagga ccctctcatc tccttggagg gtctccagac actcagggct 60gacaggcatg ggactctccc 80 785 80 DNA Rat 785 ctctcaccgt cctttggatagaggtgaaga acttggacgt ggctgtttca ggcagctgaa 60 gtcaaaagga ccactctccc 80786 80 DNA Rat 786 ctctcaccca gaagccagtt cacaaagtct tgttggcggatcttgtccat ggcgatgctg 60 taatcactga tgactctccc 80 787 80 DNA Rat 787ctctcaccgg agtatgtgga atggactgtc tgggtcttct ggacgggcag cccttggtaa 60gagacttggg acactctccc 80 788 80 DNA Rat 788 ctctcaccaa aaaggaggcggacaaacagg tttctgtaag agtgagcagg ccaaaggtga 60 gagaggagga tcactctccc 80789 80 DNA Rat 789 ctctcaccgt agatgcgttt gatggcgttg gcttcttccttatccaggat gcctttctcc 60 acataagcat caactctccc 80 790 80 DNA Rat 790ctctcacctt cgtactcgtg gacaggtggg gggaaatgtg caactgagaa cgaggactta 60attgtaagag caactctccc 80 791 80 DNA Rat 791 ctctcaccac ttcccgatgtgggggctcct ctttaggctt ggggtgcatt aaggtaaagg 60 gcagttccac agactctccc 80792 80 DNA Rat 792 ctctcaccac agcttgccac cctagggagc agggctggggtggcagagct gaagtcaaaa 60 gattaatgcc caactctccc 80 793 80 DNA Rat 793ctctcaccta aactcttcga caactatgtc ctccgagctg gtagccactg ctctctcacc 60gtgaggtggc ttactctccc 80 794 80 DNA Rat 794 ctctcaccac cgttgggtccgaacacattc agctccttga aacattctgt ttctatcatc 60 tcgttttgcc atactctccc 80795 80 DNA Rat 795 ctctcacccc ttcaggggaa ttcttgggta aggcactcgcccgaaggaat agatttccca 60 gagaaggatt ccactctccc 80 796 80 DNA Rat 796ctctcacccg agctacataa ttgcttggga tatagccttc tttcttggta gccagggaac 60gggccttcca ccactctccc 80 797 80 DNA Rat 797 ctctcaccga cctctttgggccggatctct tcagggtcag cgtaaggact ctcatatacc 60 tcggtgtcca tgactctccc 80798 80 DNA Rat 798 ctctcaccgg taatgcttaa tcatgtcact gatacagggaaaagtgatcc tcggagagat 60 gtaataacca ccactctccc 80 799 80 DNA Rat 799ctctcaccaa ccccactggc aagcagccct ctgtataggt cataaaatct ccctttccag 60gtagggctgc atactctccc 80 800 80 DNA Rat 800 ctctcaccca ggcttaaaatcgctaagatc accaaggcta gaacacagaa ggcaattcca 60 ctgaaagcat agactctccc 80801 80 DNA Rat 801 ctctcaccga tgaaatcagg agcatagaag tctcaaatatctcggctcca taatgcttgg 60 ttaaggtttc caactctccc 80 802 80 DNA Rat 802ctctcaccct gtctgcttca cagagttagc atttccagga atatcaccat tctgatgtgc 60aggagatgtg gaactctccc 80 803 80 DNA Rat 803 ctctcaccgg ttgcggcccctacgcaccac gatcaaattg gacttttcct tgggtctggg 60 gacgcagcaa gaactctccc 80804 80 DNA Rat 804 ctctcacctt acacatgatc tgtcaccaag gacaccacacgacttgaaaa gaactcggct 60 attctaattg ctactctccc 80 805 80 DNA Rat 805ctctcaccta aataccactg ctcgtgtgat ttctcatcta cgggaaaggg ggtctacatc 60aacagcctct tgactctccc 80 806 80 DNA Rat 806 ctctcaccgc ggtcatagcccttccattcc agaaccgtct accagagctg gggctagcag 60 ggatagataa agactctccc 80807 80 DNA Rat 807 ctctcaccga gcctgttcaa cttcaatcct cttgtgagggttaaatgtca acattttatc 60 cagtaaatcc agactctccc 80 808 80 DNA Rat 808ctctcaccta gatacaagtg catatgccgc ggacgactat gggtgggtta ggtttcgccc 60gcatactttt gtactctccc 80 809 80 DNA Rat 809 ctctcaccat gggactggacctctctctct ctctgctttg ggatttagat ccctgtcaat 60 acctgatgct acactctccc 80810 80 DNA Rat 810 ctctcacctc gctcacagcg aggacagggc gcagagggggcaatgctggc tgtggaatgt 60 ccccctgcag ggactctccc 80 811 80 DNA Rat 811ctctcaccag gtgcgtggat aggactgctg ggaagtcttc acacgcccgg cccagggctt 60ccagaagaca ggactctccc 80 812 80 DNA Rat 812 ctctcaccca catctgaacatagacaagca tgggagaggc ctgaaagcaa catggcatgc 60 cactgggcag agactctccc 80813 80 DNA Rat 813 ctctcacccc acagcattac aggttaaagg gtctcacatcctgcggcaag gagtaccgac 60 tggcccctac ttactctccc 80 814 80 DNA Rat 814ctctcaccga agcagaaaat agaagcaacg aggtcttgaa tatatgagac tctgcctctc 60tttgttttct gtactctccc 80 815 80 DNA Rat 815 ctctcaccaa aaactttgaccaggaacatc agcatctgac tttaaaaatt acaaatactg 60 aagcattttg gtactctccc 80816 80 DNA Rat 816 ctctcaccaa tactgcacac acatcagcac cttggcctgaggctgcaggt ccagccagaa 60 ctcagccttg ccactctccc 80 817 80 DNA Rat 817ctctcacctt ggattttgct ggacatctca atgacagcct tcactaggcc cgtcacattt 60tcatatacct tgactctccc 80 818 80 DNA Rat 818 ctctcacctt tgaattcttcctggttgatc tgcttcacga ttgcttcatc cacaagtgta 60 agtattggct ctactctccc 80819 80 DNA Rat 819 ctctcacctc tagaacccca cacggagtgg ggtttctgggaagagttgga acagcatcca 60 gatgttgggg gtactctccc 80 820 80 DNA Rat 820ctctcaccat gtttccgaga tgaagaaata taagcaattc catcagtttc ttctgaagga 60aggtctacac caactctccc 80 821 80 DNA Rat 821 ctctcaccag gaaaggagtgcggctgtcgg gggtcccaga tgccccgact gaggggggag 60 ggctccaagg gaactctccc 80822 80 DNA Rat 822 ctctcaccaa ctggcagaac tctatgatga gcactttgggtattatatgg tgtactaaaa 60 aattagacat tcactctccc 80 823 80 DNA Rat 823ctctcacctg catttgacaa gcttgaagtt cttccctggg ttgaagctgt tgctgtagaa 60gcagaccttg agactctccc 80 824 80 DNA Rat 824 ctctcacccg gtaagcatcgcgcagctctt ttttccattg caccagctca gggtcactat 60 cacactgcag gaactctccc 80825 80 DNA Rat 825 ctctcacctt cgagtgtgga taggccagcc ccaaggatgtcctagttcct gaagggccac 60 ccggtgttgt ggactctccc 80 826 80 DNA Rat 826ctctcaccaa tacagggtgg cttgatgggg actgttgggg accccgaacg atgaaagtga 60atattctgcc acactctccc 80 827 80 DNA Rat 827 ctctcacctg cggctggggcgccagggtcc tggggattgg gggtggggta ggggaaggat 60 caggaacggg aaactctccc 80828 80 DNA Rat 828 ctctcacctt tggggcacag gccctcacac ttatggcagaaaatgctgct gccattacgg 60 gtgaagcctg gaactctccc 80 829 80 DNA Rat 829ctctcaccgt gacatcaaat ctatacttga gggatgacca ccactagtct gactctcggg 60tgagacagac agactctccc 80 830 80 DNA Rat 830 ctctcaccca ctgccagcacatgactcctg cttatgtggg tgctggaggg tgggaaaggg 60 gagccagcac aaactctccc 80831 80 DNA Rat 831 ctctcacctg atgtcgatga tttttactgc gtattcttggcacgtaggtt tgtgaatgca 60 tctcctgacc acactctccc 80 832 80 DNA Rat 832ctctcacccc tgagccttct gtggccgtag gtcgtcatcg gcagtgtcat agtcctcgat 60gtccttccag agactctccc 80 833 80 DNA Rat 833 ctctcacccc tcccaaccaggcgcaagctg cattaaggag agggtggacg tcccacctct 60 gcccaggcat caactctccc 80834 80 DNA Rat 834 ctctcaccgc tgagcttggg gtctgtggaa agggcctccctctgtcgcag agcagcagcc 60 gccagcagcc ctactctccc 80 835 80 DNA Rat 835ctctcaccga gtcttggcca cacatgggac tgggtgttcc tcctgggact gcaatggcca 60aaaagccaaa tgactctccc 80 836 80 DNA Rat 836 ctctcaccga agagggggggaagcgggcgg ttgttgggga gctgggttcc gagactccgc 60 agttcatcaa agactctccc 80837 80 DNA Rat 837 ctctcacccg cagctctcac tgcagtgggg ccactggagcgcccgagcag tggaaatgga 60 cattctgcca ctactctccc 80 838 80 DNA Rat 838ctctcaccgg ggtggtgcag ggggagttct gagcctggct ccacacagca gtctcgacag 60cagcagccag ctactctccc 80 839 80 DNA Rat 839 ctctcacctc actgagaggaggatgtgctc ttcggaaagc ttccggaaag cttccgcgtc 60 aacaagccgc gtactctccc 80840 80 DNA Rat 840 ctctcaccgg ccttgaggga agagtagccg ggggaaggggctgcgagagg cgccagctgg 60 accctctgca tgactctccc 80 841 80 DNA Rat 841ctctcaccca acttcttctc ctgcagcaat gtccgtaccg agatagatgt ctccgaagga 60gccgctgccg atactctccc 80 842 80 DNA Rat 842 ctctcaccaa ctatattacagatgcatgtg ccagtatgag acatgcaagt gtgctgtata 60 cagcagatgg gcactctccc 80843 80 DNA Rat 843 ctctcaccct tgaaggcatc catgctcggc ttcttggagtgggaggctcc caagtcgcag 60 tcctcgatgt tgactctccc 80 844 80 DNA Rat 844ctctcaccga gcactagatc aggaaatggg agaagcaagg aaaggagaag aggacatgag 60gggtcggagt tcactctccc 80 845 80 DNA Rat 845 ctctcaccgt gactctaggaggctaacatt cccaccagca aacgctacta ttccacggta 60 agagtggtcg gcactctccc 80846 80 DNA Rat 846 ctctcaccga gactggaagt aggggtgact tagggccgcctctgctgaca tgcgactctt 60 ggattcatac agactctccc 80 847 80 DNA Rat 847ctctcaccga tggggtcctg agaagattat cgggaaggag acttatttct ttcattatat 60ttgccaatct aaactctccc 80 848 80 DNA Rat 848 ctctcaccgg ccaaccaaggcctttattct cttctgagcg ggtcatccag caccccccgg 60 tgtatgggga ggactctccc 80849 80 DNA Rat 849 ctctcacctg aatcagcaat gttctcagga agttctagctccttcctgct cagcagcctc 60 ttccggtcaa agactctccc 80 850 80 DNA Rat 850ctctcaccag tggccactgt ggggatctcg ggctttgtac accgtcccat aggcaccgac 60accaatttca gcactctccc 80 851 80 DNA Rat 851 ctctcaccgt ggcattgagtttgggcacga cattcaccaa ggatgttgta gctgggtaca 60 ttgggtaggg ctactctccc 80852 80 DNA Rat 852 ctctcaccgg tacaagtaac tcttaacgag cgaagaatccatgaactggc caggagggat 60 agagtccagg taactctccc 80 853 80 DNA Rat 853ctctcaccaa agtgctccca gttggatata caagtataat tcacaataga aaaaaagaaa 60cgttaagatt gaactctccc 80 854 80 DNA Rat 854 ctctcacccc ctacctctcaaagggtggga gtgggacttc attgcacaaa atactgtggg 60 agtgccacac tcactctccc 80855 80 DNA Rat 855 ctctcaccga attgtctgaa aagaagtctt ccgattggctgttggtcaca aagactgcta 60 aaatcctttt cgactctccc 80 856 80 DNA Rat 856ctctcaccgt cagtcaaccc cggcgagctt ctgaacacgg gtcctggaaa ctttctcgtc 60tattcttgac atactctccc 80 857 80 DNA Rat 857 ctctcaccaa catccaacagtaattcagtt caggaaactt gaaaggaaat cttagtgtta 60 tgacaacata ctactctccc 80858 80 DNA Rat 858 ctctcaccct cgctgaaata ttacaaccct accccccttgtcccccgtcg ctagtaactc 60 tccagtgtgg ttactctccc 80 859 80 DNA Rat 859ctctcaccgg ccgtatcgca acccgttccc ctggccctgg gcattatgct tttccatctt 60tggcttgtct aaactctccc 80 860 80 DNA Rat 860 ctctcaccgc actatggttgccagtcccgt ggttctcagt ggtatgtgcg gtgttcaggg 60 aattgaaacc atactctccc 80861 80 DNA Rat 861 ctctcacccc agaatgatta aactctactg tagaaattatatctgcttct gctacgtcat 60 catcaactgc tcactctccc 80 862 80 DNA Rat 862ctctcaccaa acacgtggct ccaagataac acatgttgcc aaagagtcat gcatgccctg 60ctcatgggct ctactctccc 80 863 80 DNA Rat 863 ctctcaccgt gacgctcaccacaaagtcaa atcagagtga tggcaggaca gtattggaag 60 ctagtcctag gtactctccc 80864 80 DNA Rat 864 ctctcaccta gacgccgcag accaaatcct aacctgggcagtggcagcgc ccaccacgct 60 catgcctcac agactctccc 80 865 80 DNA Rat 865ctctcaccat cacgcccact gcggtagagc cgctcctgtc catcccgttc ctcaggtctg 60aaaagttacg caactctccc 80 866 80 DNA Rat 866 ctctcacctg taaacagacaacttgtgagg caatctacag ggttagcaag cctcacttta 60 gtttccggag tgactctccc 80867 80 DNA Rat 867 ctctcacccc ggttaatctc ctcttccagc ttgtccacaggcagtggtgg gtactttctg 60 ttggtgctcg ggactctccc 80 868 80 DNA Rat 868ctctcaccag gctgctcctg gcatcccaac ttccaagtgt gaactagcct ggtccctccc 60ggaaggcagc tgactctccc 80 869 80 DNA Rat 869 ctctcaccgt ggaggcggtgtttcgaatct caggagtgca ttggccttga gggtccacaa 60 caacattcag caactctccc 80870 80 DNA Rat 870 ctctcaccaa ctggttcatt ctaaagtggt cacggctgttgatgacaaga ggctttgtat 60 tttatatggc acactctccc 80 871 80 DNA Rat 871ctctcaccac tgaagcacga ttcatgaggg ccacgggttc tttctgtcat atgccagcag 60aggtgccttc taactctccc 80 872 80 DNA Rat 872 ctctcaccga attacagctgctggagacgg ggagccaggg ggaggccacc tgggctcggc 60 gcgctgcaat gtactctccc 80873 80 DNA Rat 873 ctctcaccag gataaaacta tcttgggggc tcagagcagggtgcaaaaga gtcgtttgtc 60 gcggaaggga ttactctccc 80 874 80 DNA Rat 874ctctcaccgt ttgtgatctg agaaagagtc gcatctgcac atgctttacg ggcatctcta 60atttggttct tcactctccc 80 875 80 DNA Rat 875 ctctcacctg gcgtcccggatctggttccg gagctgctca gcctcctgtc tcagttgctc 60 cagctcactc atactctccc 80876 80 DNA Rat 876 ctctcacctt atgaagggag aagctatgag gactgtggagaggagaggcc ctggaggaaa 60 tggggcagtt ctactctccc 80 877 80 DNA Rat 877ctctcaccca cagacctctt taataaaata atcattactc aaataattta aaaatctaca 60aaagacacaa gaactctccc 80 878 80 DNA Rat 878 ctctcaccca caatgacatatgcatccccc atggccatgc agtggccagg cagccagcag 60 cctccatcct gtactctccc 80879 80 DNA Rat 879 ctctcaccta gcagtctgcg tctctgactg acagctatcctgcgtgtgtc tgtacccaag 60 agcagtttat caactctccc 80 880 80 DNA Rat 880ctctcaccgg cctccaagcg gcagagagtg gggtaggcag gaggctccct gtaaacattt 60ggacttgggt caactctccc 80 881 80 DNA Rat 881 ctctcacctg tcaggatctgcatgtctgag ttggtgaagc tgcaggcaga caggtagttg 60 gtgtgcatag cgactctccc 80882 80 DNA Rat 882 ctctcaccct cactgtctgg agtttactgg ctgctaggatgcagtcccca agcctcgggg 60 atccttccat ctactctccc 80 883 80 DNA Rat 883ctctcaccgg gtgtcgaggt gagggtggcc ggggtctggg gctccggagc cgagagagga 60ggcgcccgcg ggactctccc 80 884 80 DNA Rat 884 ctctcacccg aaaggtccttctattaaaga cacgatacag aacaactttg gtcagaactc 60 tggtcaggtc caactctccc 80885 80 DNA Rat 885 ctctcaccga gcacagcatg cttggatatg ccattggtggtcttttatag aagtgagctt 60 caaaaactgg gaactctccc 80 886 80 DNA Rat 886ctctcaccca ggaaaccttg accgccggag agcgttgccg tctccagacg tgcgtgcgcg 60tgtgcgtgtg cgactctccc 80 887 80 DNA Rat 887 ctctcaccac agtccaagactgtgaggatg aaggtgctta tgtgatgtcc ctctgccgtg 60 gtggccggtg aaactctccc 80888 80 DNA Rat 888 ctctcaccgg cgcggccggg ctgcggggcg gcggggacggggcgggcggc ctcgggcggg 60 ccgggaaagc ggactctccc 80 889 80 DNA Rat 889ctctcacctc tctccttgta atatcataca ctagtaaagc ccccgctgca cctctgtaat 60atgaccttgt gaactctccc 80 890 80 DNA Rat 890 ctctcaccct ctaaactgagacaagctggc gacacggacc gtggcgactg cgctggcgac 60 cgggccccag aaactctccc 80891 80 DNA Rat 891 ctctcacccc aagaacttaa acaagaaatc gtaagtctcggacatggcgg tctcgccatc 60 ctgcgccaca gcactctccc 80 892 80 DNA Rat 892ctctcacctt aagacatggg aagattctta agtgtaagga agtaataaaa cacagaaagt 60gaccacaaga atactctccc 80 893 80 DNA Rat 893 ctctcaccct gtgcaaacttttctccttgc tgcctggaga tttctctgtc tgtttcacag 60 tccagcttat ttactctccc 80894 80 DNA Rat 894 ctctcaccgc tgctaatgtc gtagactaaa aatattccctgggctcgccg gtagtactgt 60 tttgtgatag tcactctccc 80 895 80 DNA Rat 895ctctcaccac tggttagccg tcctccctgc ggggctgagg gtttgtgttg tacaccagac 60tcggcagcat tcactctccc 80 896 80 DNA Rat 896 ctctcaccct tggcagcggccagttcatag gcagatttca aggccaaact ggtggacctg 60 agatggagag gcactctccc 80897 80 DNA Rat 897 ctctcaccca acaaaccatc tccccgaagc tgtggccacgggcatgtaca gccacgccca 60 gctttttaca tgactctccc 80 898 80 DNA Rat 898ctctcaccta acacagactt gcaacagcgg ggccaggcag gacgttgggg ccaattggcc 60ctggtctcaa gcactctccc 80 899 80 DNA Rat 899 ctctcaccag ctggccaatccggtctaagt tatccaggaa gtacttcact gactcaccca 60 gctggaactc gcactctccc 80900 80 DNA Rat 900 ctctcaccga tccacagagg tgaataagag agcacaggctcccctgggca agttagtcct 60 ccaggtcctc agactctccc 80 901 80 DNA Rat 901ctctcaccgg tctagcacct gctgagtggt tgtgaaggct ctgtaggtgc acaggaatac 60ggtgacgtag gaactctccc 80 902 80 DNA Rat 902 ctctcaccaa aacaagttgggtcttacccc aatgaaaacc cccacaccac agctaagtct 60 agttttacat ggactctccc 80903 80 DNA Rat 903 ctctcaccgt ggtagcagtt tctcagggcc cagctctgcactcagcatgg cccgtgcccg 60 actcagggcc tgactctccc 80 904 80 DNA Rat 904ctctcaccac agcccacagc tccccctggt actcaagaag gaactgctgg aactcagaaa 60gggacacctg gcactctccc 80 905 80 DNA Rat 905 ctctcaccgc aggtagacggctgaggcatc aggccggtct gtgttcccta ggatgaacac 60 ggaggagtct ttactctccc 80906 80 DNA Rat 906 ctctcacctg gatggagatc ttcacaaaaa gtgtggctgatggatgctgg tctccattct 60 tagacaagag gtactctccc 80 907 80 DNA Rat 907ctctcaccgc cccgaggagc cctaacactc ggatggcctc tctccgagtg ccctggttct 60gctccgtctt caactctccc 80 908 80 DNA Rat 908 ctctcacctc ttcttgcatgcaggtgtaca caaaatggga ggctggatgg aggctagtgc 60 cgtctttgga gaactctccc 80909 80 DNA Rat 909 ctctcaccca gtctttcaaa tcgataggtg ggggcgaaggtgatctcctc ctcctcaaag 60 tgcaggaaga ctactctccc 80 910 80 DNA Rat 910ctctcaccac cacgtagtgt tcgcgccttc ctgtgagaac agggaggaca tcattggatg 60tggcttccgt taactctccc 80 911 80 DNA Rat 911 ctctcacccc tctctaggattctggagggt agggtgtaac cctgggctct gtccagccca 60 gtggtacaca gaactctccc 80912 80 DNA Rat 912 ctctcaccaa aacaaagctt ccatatctga gagtgagtgctcccggcttc cgttctgaac 60 ccaacagctg tgactctccc 80 913 80 DNA Rat 913ctctcaccgt ctcagcactg tggcgagtgg cttctctcta gttacggagc tctttttgac 60cttctttggc tcactctccc 80 914 80 DNA Rat 914 ctctcaccca tagtaacggttgaccaaaat ttgccggagt gcctcacccc ttttatggtc 60 ttccagttgc ctactctccc 80915 80 DNA Rat 915 ctctcaccga gccctggcca ccaacagaat gaactctttccacccttcca gggggttctc 60 taatgtgggt ccactctccc 80 916 80 DNA Rat 916ctctcaccgt tttcaggaat ccgggcagga acattagttg tgtcgaagtt ttcacattca 60agaacttcca caactctccc 80 917 80 DNA Rat 917 ctctcacctg acagcagcagcgtcctacgc ttcatgtagt acttgtgcag ctgggaacct 60 cggttggttt tcactctccc 80918 80 DNA Rat 918 ctctcaccga gcctgtttca gatgctccag cctccgttctgtctctgcat ctgcctgggc 60 ctccctcagc tcactctccc 80 919 80 DNA Rat 919ctctcaccaa taaatttgcc tccttcctca tatgctgcta tccttaaaca ggccagagaa 60ggcagaacta ccactctccc 80 920 80 DNA Rat 920 ctctcaccct cctttgcccggaatgggttt gagaacaagt tggtgacact actgagggtg 60 ttaacgaggc gtactctccc 80921 80 DNA Rat 921 ctctcacccc gcgttgctgg atgcggatcg gggtcctcggctgccgccac aggtgctggg 60 ggggcggctt caactctccc 80 922 80 DNA Rat 922ctctcacccc accgctgcga tcccaccgtg agtggtgctc actggtcagc aatcctgacg 60ctcctgagct ccactctccc 80 923 80 DNA Rat 923 ctctcaccgt catgggatccaagggactcc gtcccgctga tgtgccattg tccacatcga 60 aacatatcca ggactctccc 80924 80 DNA Rat 924 ctctcaccct cagaggacac gacaggctgg cactgtggtgcagggagcca gggtgtagac 60 aaccccctcc caactctccc 80 925 80 DNA Rat 925ctctcacccc caggaagtat taaaagccac tgacgacaca ttgggagcca ggaaagggca 60gtctgacgac acactctccc 80 926 80 DNA Rat 926 ctctcaccga aggccgcttggcagcgatcc cctctttcct aggctgctcg attgcaggag 60 ccctgagcag ggactctccc 80927 80 DNA Rat 927 ctctcacccg ttggtgtgct ccaggaaggc tggtacgctccttgttgacc cctggatcac 60 tgactgtgga ctactctccc 80 928 80 DNA Rat 928ctctcaccgg tgtcttcaag ctccttagcc agctgctcct cctggtctgt ctggacccca 60aagcgcggga tgactctccc 80 929 80 DNA Rat 929 ctctcacctc gccaagctcgccggcaaggc aaggcagcac ggaactgcta tttgcttcac 60 ggccttgcat tcactctccc 80930 80 DNA Rat 930 ctctcaccgg cgatgatgcc tgatgatttc aaaagccacatgagagaggt ggaggccttt 60 ttgctgcatt gcactctccc 80 931 80 DNA Rat 931ctctcacctg aacctcatcg ttaggaggaa tgtgtttctc ttctttacca cctgggcgga 60ctcccgctcc agactctccc 80 932 80 DNA Rat 932 ctctcacccc aggcgctgctgctgttgtgg ggtcaccgct cccccagatc gcttggtgga 60 gccgcgggtc gaactctccc 80933 80 DNA Rat 933 ctctcacccc ggccatcacc atccttatcc aggaccaagatgcttttgtc caccagctgc 60 tgcagctgcc agactctccc 80 934 80 DNA Rat 934ctctcaccag gctgtgatca gagatgactg caggtgggat tctgccatgg tgcagcggct 60agtttgagag agactctccc 80 935 80 DNA Rat 935 ctctcaccgg gtgtactcctggagcttagg ttcatcatag ggacggttgg ccttcttcag 60 gaggtcggac agactctccc 80936 80 DNA Rat 936 ctctcacccc gtccttatct gcgtttatta tggttttgtctacaatctgc tgtaactgcg 60 tatctttcag gtactctccc 80 937 80 DNA Rat 937ctctcacctg ttccccacca tctggtaaat ggcatccact atgtccagca tctcgtttct 60ggtgatgtag ccactctccc 80 938 80 DNA Rat 938 ctctcaccaa aaaaatatatatatttaatt cagactaacg gctaaaccgt ctctgtgagt 60 ggttctctct cgactctccc 80939 80 DNA Rat 939 ctctcacctg agatgggggt aaggaggcgg ccgtacctccttcttgaact ggggactcgg 60 cttgaagtga aaactctccc 80 940 80 DNA Rat 940ctctcaccct ggacgcgatc cgctggcttc tcgagctcaa gttcggcggt ggctgaggct 60ggggagataa ccactctccc 80 941 80 DNA Rat 941 ctctcaccgc agctcaaaaaaaaaaagtga acaggaagtg gtcactgaat attgctgcta 60 ttactgccgt tgactctccc 80942 80 DNA Rat 942 ctctcacccg gtgatgatgg agaaaggata atgaaagacgcagttatgag ctttcatcct 60 aaaaggctct ctactctccc 80 943 80 DNA Rat 943ctctcaccct aactatgcca aacatgtgta actcatgtaa aaattccaca tcccatattg 60gccacctcaa gaactctccc 80 944 80 DNA Rat 944 ctctcaccat gagtggtataaaaattcaca gtcaatgtac ttcattttgt tttacataag 60 caatcaaagg gcactctccc 80945 80 DNA Rat 945 ctctcacctc caggagacaa actgcacact ctgcatgcaacagggcacgt ttatgggaag 60 atttatccac agactctccc 80 946 80 DNA Rat 946ctctcaccca agttgtcgtg tgtcagtttc cattagctga aagaaggggt gagagggtca 60aattcatttt tgactctccc 80 947 80 DNA Rat 947 ctctcacctt ttcccaaagctgcagtgtga aaagactata aacagttgac cccatacaca 60 tgaatgggct tcactctccc 80948 80 DNA Rat 948 ctctcaccta catccagaat gtcacaacag attaacttgagctcagtttc aaccatttgc 60 cggtactccc gaactctccc 80 949 80 DNA Rat 949ctctcaccct tgcgatgtcg tttgcggagc ttgagtagta ggactgtcaa aaagatgata 60atgagcagga agactctccc 80 950 80 DNA Rat 950 ctctcacctg gggtgcaaagccaacagaga tttcagttta tttacataga ttctctagct 60 tcttggtctt cgactctccc 80951 80 DNA Rat 951 ctctcaccca cagtaggtga ctggctgtag gtccaagttattgtgtgctg gggacatcgg 60 attcggggag agactctccc 80 952 80 DNA Rat 952ctctcaccgt atgcagtggg gagcatggga ctgtactggg aagctggagg cactggagta 60tggacgccct ggactctccc 80 953 80 DNA Rat 953 ctctcacctc gaacacatccaagtccactg aggggggatc acaatcaaga agctgtcgta 60 actctgcagg gtactctccc 80954 80 DNA Rat 954 ctctcaccct tttttttttt cggggtctcg ctgctgggagacggcgcggc cccgtcctcc 60 gccttggggg acactctccc 80 955 80 DNA Rat 955ctctcaccta ggaagatagt catgaccaca atgacgctga tcatgatcag agtgttcagc 60acggagttga ggactctccc 80 956 80 DNA Rat 956 ctctcaccaa agttactctggggtcggcca ttagatatcg actcagggtt gtcaagtctc 60 tgtctgtcat gaactctccc 80957 80 DNA Rat 957 ctctcaccca gagtggatct gagaaagggc aggagagacagggttcaggt gggggcaacg 60 tggaaagccg ttactctccc 80 958 80 DNA Rat 958ctctcaccga gggcagccat cagcaggact actagcaaag cacacgccac acccacaccg 60atgaagacgt tcactctccc 80 959 80 DNA Rat 959 ctctcacccc tatagttatgagcagaattc actgtaagtc acacgagtca tcctcagtgg 60 tcaacgtctg caactctccc 80960 80 DNA Rat 960 ctctcacctg cccagtttgt gtttgtttct ctcacccatcagtcagtcca cctcctctgt 60 cttcggaagt ccactctccc 80 961 80 DNA Rat 961ctctcaccat gagaggaaag ctgagggcgg agaagaggaa gtgaggggga aggaggggtc 60tggttcactt ctactctccc 80 962 80 DNA Rat 962 ctctcaccgc ctcaggggactgagacgtga tgccagacac aagagctacc tcacttcact 60 ccccacaacc ttactctccc 80963 80 DNA Rat 963 ctctcaccag ggcccaggta ggggcagtgg taatctggttttagcctggt ccctggtatg 60 gagggcttga tcactctccc 80 964 80 DNA Rat 964ctctcaccca taaaaaagat aatataaagg aagacaattt gctgagtatt aaaagttatc 60tcaatatgaa ctactctccc 80 965 80 DNA Rat 965 ctctcaccct ttgatagctcatgggacagc cagtccagac catcatacag gcctgtgcct 60 tgggtggcac agactctccc 80966 80 DNA Rat 966 ctctcaccaa gaaaaccccc aaccccacta aacggtagaacagtaactcg agttcagttt 60 caaaaaagag agactctccc 80 967 80 DNA Rat 967ctctcaccaa acagagcgaa gagctaccag atacagacag gcttggtcca cactcgctgc 60tgcttgtcaa gcactctccc 80 968 80 DNA Rat 968 ctctcacctg tggactccccaagttctaac tctgataaga cgtcaatctg tagggagggg 60 tccacaccaa gtactctccc 80969 80 DNA Rat 969 ctctcacctt cccggcaggt cttggttaaa atccgctggccctcagcaag aacttttcct 60 ccatagatac atactctccc 80 970 80 DNA Rat 970ctctcaccct ctgctgcttg agagcttcaa agtaagggat gattttcgtc ttccctcgac 60tcttatcaat gaactctccc 80 971 80 DNA Rat 971 ctctcaccgg gccgtcggcttttctgaagc attaccacaa atcagggaca gtatcttttc 60 cagaagtacc aaactctccc 80972 80 DNA Rat 972 ctctcaccct gggtcacccg gatggtcttg ggggttccatcccctgtgaa ggtggtctcc 60 agatgtgtag tgactctccc 80 973 80 DNA Rat 973ctctcacccc tgtttttgaa tcaagttttc aatgatggct ttggtctcag gttgggaagc 60attacttgtg aaactctccc 80 974 80 DNA Rat 974 ctctcacctg tggtccaggtggtcagccac ttgggacaag gtggcgctgt ctgagcagga 60 cacgaactgg ttactctccc 80975 80 DNA Rat 975 ctctcaccgc cccatcagca cttgggcctc tagaagaccatgtatattga tttgcaggta 60 gtttctctct caactctccc 80 976 80 DNA Rat 976ctctcaccca cagtctcaga gaactgggca ggagtggcca gcttcagcag agtgatgtca 60ttgcgcacag tgactctccc 80 977 80 DNA Rat 977 ctctcaccgt ggcatagtccacaaccagca gacggccctg ctgcaggacg tcaggagtgg 60 ctccgttggt ctactctccc 80978 80 DNA Rat 978 ctctcaccca caacattcac catcttttcc actcctctcctgtcattctg gacagtattg 60 taggacgtgt atactctccc 80 979 80 DNA Rat 979ctctcaccaa cggagcatca ccaaacctcg ggggcaggca gatggtctgt atggtcctgg 60atggctgtgc gcactctccc 80 980 80 DNA Rat 980 ctctcacctg gcatcacagagggtcaggat ggacacattc tgggcccggt tcagccaggt 60 cacagcaacc ttactctccc 80981 80 DNA Rat 981 ctctcaccca ggggagacag agtacaaaga aggccagggttccatctggc caggaagcag 60 tgagttaaag acactctccc 80 982 80 DNA Rat 982ctctcaccgt cagaccttca cccttaaaac gtacaagttt gtgaagtacc tgtcacaaag 60taaactctgt ctactctccc 80 983 80 DNA Rat 983 ctctcacccc cccttaaataagggaacaag atttggacca gcctagtaag ggggaaaaaa 60 attgaaattc tgactctccc 80984 80 DNA Rat 984 ctctcaccca cattcaagtc tagcccagga gtcccgtcagcttgcccagt gttgaacagg 60 taccattgct ttactctccc 80 985 80 DNA Rat 985ctctcacccc cagcctgtca cccgcccttt ataaccagcc tggagcaaac tggttactgt 60ctgcttgtct ggactctccc 80 986 80 DNA Rat 986 ctctcaccgg atagtgaaccggtgatttac gaaaggtaag agaactcagt tgttcgccat 60 agtctcctga atactctccc 80987 80 DNA Rat 987 ctctcacctc catccagctt aagcagggcc aggtcgttctcgtatttgag gttgtagcca 60 gggtgtttaa tgactctccc 80 988 80 DNA Rat 988ctctcacccc aggagcttga ggtctctagt ggcttggcca ctctgcttca gcgcaacact 60cagatgtcgg agactctccc 80 989 80 DNA Rat 989 ctctcaccag aaaacctgcacgattccagc tccagatttc ctcacacact ggcccgagat 60 ccagagttaa ttactctccc 80990 80 DNA Rat 990 ctctcaccgt ggggcagtag agcttggtgc catccccaggctgcctgagg gtctggtcag 60 agcaggtttc tgactctccc 80 991 80 DNA Rat 991ctctcaccac agttcacaac gtaattattt gctctcttct ctttgactcc cagggcttgc 60atgatcaact gcactctccc 80 992 80 DNA Rat 992 ctctcacctg tagttctccttggtgatgct tccgtcaaag accctccagc gccactggtc 60 aatgaggtag ctactctccc 80993 80 DNA Rat 993 ctctcaccat gtctgcagtc ccccaactaa ccctcttgaagttcagtggg atttgcatac 60 tccacattct gaactctccc 80 994 80 DNA Rat 994ctctcaccca ggaactgtat tcgtgagcag taccactccg cgtctcatca tacgggtaat 60tagccacgag gtactctccc 80 995 80 DNA Rat 995 ctctcaccat gggctggctcatctggaaga agaacttatt tggtcctctc ctctcccatg 60 tgcttgcact caactctccc 80996 80 DNA Rat 996 ctctcacctg gctgtgttgt ccagtttgat gacaatgtctacatcatctg gtgggctgag 60 accttcttta tcactctccc 80 997 80 DNA Rat 997ctctcaccag tcccaaacta gagtttgtcc aaccacgttg ttggcgatgc tgacgatggt 60ggaggtagca tcactctccc 80 998 80 DNA Rat 998 ctctcaccag catcgctgtgatccccccat tcccagtgct agtgttgaag tgcataaaga 60 agccagagtc ttactctccc 80999 80 DNA Rat 999 ctctcaccat accggaggcg ttcttgtccg gtactggatgcctagggctg ccaggcaggc 60 caccagccct gcactctccc 80 1000 80 DNA Rat 1000ctctcaccta tgtagctccc attttgctct gccatagcac atgctagagt aaggaaacca 60cttcagtgcg ccactctccc 80 1001 80 DNA Rat 1001 ctctcaccgc agtgcaatctcattggtatc tgtcccagcc tgggagctca aggttggggt 60 tggggaggtg ggactctccc 801002 80 DNA Rat 1002 ctctcaccca ggtcctagta atccctttgt atatctaaagttctggatgc acacagagcc 60 tccagccacc cgactctccc 80 1003 80 DNA Rat 1003ctctcacctg tgattgtgca ggtaaagaat gatggcatgc tgaaacacgt cttcactaag 60gtaacttttc aaactctccc 80 1004 80 DNA Rat 1004 ctctcaccgg atccggaagtagccactctc accccattga gagccccagc tgttcttgac 60 aatccagtag tcactctccc 801005 80 DNA Rat 1005 ctctcaccca tcctgcccca catacgggta agcgtcttcagagtcaatgc ctccattctg 60 ctgcacatat tgactctccc 80 1006 80 DNA Rat 1006ctctcacccc attgcagcct catcattgag tgtgatgttg acaacattct tgacgaacgc 60gacggccttt tcactctccc 80 1007 80 DNA Rat 1007 ctctcacccc catagaatgttccagattgt gaagcattat aaacttgaga ttcttttccc 60 agatgagacg tcactctccc 801008 80 DNA Rat 1008 ctctcaccac gggtttggtc agttttaaga tgaactccttcggatgtagt gtccgtcagt 60 ccttattatc gcactctccc 80 1009 80 DNA Rat 1009ctctcaccgt cagacatggc ttccactgcc ccaaatgccc aacaagagcc acaggacccc 60tggtctctga tcactctccc 80 1010 80 DNA Rat 1010 ctctcaccag tttcagcatggcgcaaagtg actggatgaa ccatgacccg tcccttgaat 60 ttctccagga atactctccc 801011 80 DNA Rat 1011 ctctcacctt gtgtcaacct ttaaaatatc tgcaacttcattagagtatg tgatcccaca 60 tattccctcc tgactctccc 80 1012 80 DNA Rat 1012ctctcacctt acagcttcca ttgcatattc gatttgatga atcctgccct gagggctcca 60aacagtgaca tcactctccc 80 1013 80 DNA Rat 1013 ctctcaccag gatggatttctgctttttct cagtggctaa taccacgcca tttgcagctt 60 taattcccac tgactctccc 801014 80 DNA Rat 1014 ctctcaccat ggtgaaatgt tacatattgt cgtcatctgattcatcttct tccttcaaag 60 attccttggc atactctccc 80 1015 80 DNA Rat 1015ctctcaccgg ctctggcact ctaccctggc tctgttgatg actatccttg catcggcagt 60gagacctgag acactctccc 80 1016 80 DNA Rat 1016 ctctcaccca actgtcattgtctcattata ggtgaaccag tggttctgtg tctccactct 60 ggctttatca atactctccc 801017 80 DNA Rat 1017 ctctcacccg tccagctcat gcaccagctg ccgataatcacccacatgag gctgcttggc 60 tgctttggcc acactctccc 80 1018 80 DNA Rat 1018ctctcaccgt tgggctccca gagagactca cacatcccat acatttgttc agagcaggtg 60ccactgacta cgactctccc 80 1019 80 DNA Rat 1019 ctctcaccga aggtaatgttctccagattg tggatgccat ctttgtcaat gacccgaacg 60 ctgaaggtgg gcactctccc 801020 80 DNA Rat 1020 ctctcaccct cttgtcattc acaatcaact ggagttcttcaatcttggac agataatatt 60 gacggagtcc acactctccc 80 1021 80 DNA Rat 1021ctctcacccg tcactgtatg tgacatcagg tttttcctcc acctgcatca tggtaactgt 60tgggtcaatc ttactctccc 80 1022 80 DNA Rat 1022 ctctcaccga ggaactccttcttcaggttc ttctgctcat ccttgatata ctcctcctgc 60 acctccagga acactctccc 801023 80 DNA Rat 1023 ctctcacctc tacaaatgac aggatgctga cgtagtgttctgagcccacc gatgtggaca 60 caatggcgtg atactctccc 80 1024 80 DNA Rat 1024ctctcaccgg ggcttcataa gctacaccaa gcatgtccac ataaccaaga aagctttctc 60caccagcata gcactctccc 80 1025 80 DNA Rat 1025 ctctcaccca atacagtacctccgttgaag gcataaggcg agaaacgcat ctgcacaggg 60 ccggcggagc ccactctccc 801026 80 DNA Rat 1026 ctctcaccaa gacttcatca agaagcttgt ggatgttgcgcctctctgct gcaagcagaa 60 cgccatcatt ggactctccc 80 1027 80 DNA Rat 1027ctctcaccga tctgtccaca agcaatgact gtcactctcc agtttgcaag ggatggctga 60acagggaaac acactctccc 80 1028 80 DNA Rat 1028 ctctcaccta gcatgggatcatagggcagc gtgtgatctt gcactcgcag cccatctggt 60 acctgtggtt taactctccc 801029 80 DNA Rat 1029 ctctcacccc tctgtgattc cagagagatc agcatcattgttgaagaccc gggtgatgcc 60 cagtgagctc agactctccc 80 1030 80 DNA Rat 1030ctctcaccct gcagtagcca cccttctgcc ggatgcaggc gtagtgtttg gactgatagc 60cagggtaccc gaactctccc 80 1031 80 DNA Rat 1031 ctctcaccag gacagtcatactcatggtta aggggcatgt tgacggctgt ctccgagttg 60 gggtcttgtt taactctccc 801032 80 DNA Rat 1032 ctctcaccga agctcaggtg cactttgttg cgaagacatttttcaacttg aaactttaca 60 gaatctgcaa tcactctccc 80 1033 80 DNA Rat 1033ctctcaccca tgcgggctga gactagaatg gctgtggagg aagacgccac tgtgccgctc 60tcgttcacct cgactctccc 80 1034 80 DNA Rat 1034 ctctcacctt ttcttgatatgtaggttgga caaaatctgc tgaaagctcc ccaatcagct 60 ctgactcact caactctccc 801035 80 DNA Rat 1035 ctctcaccaa tgtagaccaa aacatcatct tcatccagtgtctctttgct gatccacttg 60 ttgaagttat cgactctccc 80 1036 80 DNA Rat 1036ctctcaccgg gcactttctc ttgcttaggt tactcaggat ctggtcctgg ttgggcctgt 60agagcaccac gaactctccc 80 1037 80 DNA Rat 1037 ctctcacccg cccacacggtcgtacacaca cttggtctgc tcgatgagct gtgtggttag 60 agctctgatc tgactctccc 801038 80 DNA Rat 1038 ctctcaccgc tcctggctct gcctgaaatc atccagccccaccacgccgt ctgccagttg 60 gcccagacca ccactctccc 80 1039 80 DNA Rat 1039ctctcacctc ccaatgaaac atcttcatga ttgagaaccc atttcttatc ctttggaata 60ggattcagca gaactctccc 80 1040 80 DNA Rat 1040 ctctcaccca gggctctggccagagggctg ggcagaaaca agaggtaggc ctccacgctt 60 gcacggtggc tgactctccc 801041 80 DNA Rat 1041 ctctcaccga gttcatgctg gctgttgatg tcacggtagaccacggtata gttggtgatg 60 cgaccattcc tcactctccc 80 1042 80 DNA Rat 1042ctctcacctg ttgtcaactg cagctttcgt gacatctgcc gttgtcatgt tttggatgtt 60ggcactcact ttactctccc 80 1043 80 DNA Rat 1043 ctctcaccaa ctggccaccacgagacttcc gaagtgggac catcacaatg aagtagttct 60 gcactgtcac ggactctccc 801044 80 DNA Rat 1044 ctctcaccaa ggtcaccaaa ggagtgtcca gagtgagtccccaaaatggc cttggagagg 60 cttctgctga agactctccc 80 1045 80 DNA Rat 1045ctctcaccca tggtgacgta ggtgatttgc aaccacaagg cactcgtcgt ggtattgttg 60gctttcatgc tgactctccc 80 1046 80 DNA Rat 1046 ctctcaccac ggccacatcagctaccgcca gtgacacgat gaagcagaag gtggcatcgc 60 gaagtgcctg gtactctccc 801047 80 DNA Rat 1047 ctctcaccca gacccagcag gcaggggagg ggggaagtcaataaaataga atgggtttcc 60 tcacattttt atactctccc 80 1048 80 DNA Rat 1048ctctcaccga agtcccggtt cctgtaggca tagacaatgg gattgacaac tgaattggcg 60tgtgacagga ggactctccc 80 1049 80 DNA Rat 1049 ctctcacctc ttggaggtggcaattaaaag tttcttccca aaactctttg gcaaaaccat 60 tgtggacaga ctactctccc 801050 80 DNA Rat 1050 ctctcacccc acttcctctg cacggtacag agctttctccgtccttcaat ggacatggtt 60 tctcatcagg gtactctccc 80 1051 80 DNA Rat 1051ctctcacctc tccagggagc tggagctgcg gaggggcagg agtcggtgac cagggttccc 60acctctgcgc ccactctccc 80 1052 80 DNA Rat 1052 ctctcaccgt acatggacagcaccgtgatg atgaggcgga tagccctccg ccttttcttc 60 tccgagtgct cgactctccc 801053 80 DNA Rat 1053 ctctcaccgc tgcggtggtt ctgggaccag acggggaggagcacggagat gcagcggtcg 60 aagctgatga caactctccc 80 1054 80 DNA Rat 1054ctctcaccac ttcacccagg tcagaacgtg gtgtctcatc agggtgagct catactcctg 60gaagttgtta taactctccc 80 1055 80 DNA Rat 1055 ctctcaccct gctgactgaggcggcgaagg acgccaccag gaagcccacc atgagcaggc 60 tcacagtctc cgactctccc 801056 80 DNA Rat 1056 ctctcaccga tgttctccgg agtaccagct tctatacagtcccgaggcag cccgcttaaa 60 ttgaccttcg ggactctccc 80 1057 80 DNA Rat 1057ctctcacctc ccaggagggt gggaggaagt agttacagtg gctatcccag ttagagccgt 60agaaaaagaa gaactctccc 80 1058 80 DNA Rat 1058 ctctcaccct aatgactctgaagctcccca gctcagacct ggcagagtgt catcagatga 60 ccacactgac ggactctccc 801059 80 DNA Rat 1059 ctctcaccgg gagctcacca gtgtgtgtcc ctgtctcgcaaagtctgtgg ccgtcctgat 60 cctccgagtt ccactctccc 80 1060 80 DNA Rat 1060ctctcaccag accctcagcc ccaccctgcc gaaggctgcc aggacaaagc tggagcctgg 60gagatccgaa tcactctccc 80 1061 80 DNA Rat 1061 ctctcaccag ggggtgcaggggcaggtggg aaacacccaa ggtctaggag cactggatgg 60 acatgtaagg ccactctccc 801062 80 DNA Rat 1062 ctctcaccaa aggactcatg acaggcaggt tgaccatggtacggatcttc tgccaatggc 60 ttttctgccc caactctccc 80 1063 80 DNA Rat 1063ctctcacctg acactgaatc gcgtcattcg ggccagccct ccgaatccct gctccgaacg 60ggaagtgatt cgactctccc 80 1064 80 DNA Rat 1064 ctctcaccgt ccagcaggaagcccgaggct gcctgggcct caagtaagat aagggctgtt 60 ccagggctga gcactctccc 801065 80 DNA Rat 1065 ctctcaccgc gctccaggtt ggcgatggtc tcctggcagcggcagagaca ggccttgatg 60 gacttcttcc agactctccc 80 1066 80 DNA Rat 1066ctctcacctc acgaatcaca ctgatcacga ctttatttaa aacttagcag acaacactgt 60gcagaagcat ccactctccc 80 1067 80 DNA Rat 1067 ctctcaccta ggggtcgtccacagtctgcc ccacatctcc taccagaatc tccttgccct 60 cctccaggat gaactctccc 801068 80 DNA Rat 1068 ctctcaccaa ctgcgtgtgt gtccactgca tttccaggcatgcattccgc tcccgttgca 60 ggaagggggc agactctccc 80 1069 80 DNA Rat 1069ctctcaccaa aacaaaggaa gccctggggg tccaggcagc tggagtcagc agagagggct 60gctgtacagg gaactctccc 80 1070 80 DNA Rat 1070 ctctcacccg tactcctcgttcatagtgat cactccgtcg cacgtctcgt ccagtaccag 60 ccggcgcgtc atactctccc 801071 80 DNA Rat 1071 ctctcaccag tttctcccgc ttgagatgct catacatcaccgggatagcc attgccgccc 60 gaagctgccg ctactctccc 80 1072 80 DNA Rat 1072ctctcaccag tgctaggtaa agggtgatgg gacagtggtc actgccaaga gccttggacc 60ggatcttgct gtactctccc 80 1073 80 DNA Rat 1073 ctctcacccc cagggacagggtcctggggc tggctgggca gacgcctgct tacaggctgc 60 ataaaggcat cgactctccc 801074 80 DNA Rat 1074 ctctcacccc ttcagcagct tccataacac ctgtctggtgaatgaatctt gctggaacac 60 agggtgatgg agactctccc 80 1075 80 DNA Rat 1075ctctcaccgc tgcggtcatg tcgtctattt tgtagacaat gttggttggc gctttctctg 60catgtctgat taactctccc 80 1076 80 DNA Rat 1076 ctctcacctc gcacgggcaaagttgacgaa agagatctca tcgaagcgga tgtgcacagg 60 gggcttgtgc acactctccc 801077 80 DNA Rat 1077 ctctcaccga tgacaggagg ttgagcgcca gcatagaaggccgtggctgg taggcccacc 60 gggagaggct ctactctccc 80 1078 80 DNA Rat 1078ctctcaccga atgcaggtga ggagcatgta actgagggag gccgcgataa acacaatgaa 60agcattttca tgactctccc 80 1079 80 DNA Rat 1079 ctctcacctg gcagcgttcacgaaggcttg agaagtgtgg ttcatggtag cgatccaccg 60 cgatcgcaca caactctccc 801080 80 DNA Rat 1080 ctctcaccta acccaaataa acaaaaagtg gactacattaccacaaaaaa agtactgaat 60 atgccagaat ctactctccc 80 1081 80 DNA Rat 1081ctctcaccgg taagtaataa acttaaaaat gagaaaagtg gattggaaac tgcataaagc 60tacaagtgac tgactctccc 80 1082 80 DNA Rat 1082 ctctcaccgt catggatgaccttggccagg ggggctaagc agttggtggt gcaggatgca 60 ttgctgacaa tcactctccc 801083 80 DNA Rat 1083 ctctcaccaa tggagaactc cagcttggac ttctttccgtagtcgacaga gagcctctcc 60 atcagcaggg agactctccc 80 1084 80 DNA Rat 1084ctctcaccac taggcagatg acactcttaa gagactttgg gcaccaagtt cacagtaaac 60acttgagatg ggactctccc 80 1085 80 DNA Rat 1085 ctctcacctg tgaattgatccctgtatttt ttacacctcc acaagtggaa gatccttttt 60 agctccttgt gaactctccc 801086 80 DNA Rat 1086 ctctcaccgc caggatagag ccaccaatcc acacagagtacttgcgctca ggaggagcaa 60 tgatcttgat ctactctccc 80 1087 80 DNA Rat 1087ctctcaccac tccagtagag ctgctggtga cccatcttgc cttcagacgc ccaaggagga 60aaaaggaacg agactctccc 80 1088 80 DNA Rat 1088 ctctcaccat gccatcagtgcccgccggga atgcagactg aagaattaat agccacccct 60 caggcggagg acactctccc 801089 80 DNA Rat 1089 ctctcaccga caggaagtgg ggtgacagcc taacagtgtttcttggtgtc aaggtctttg 60 tattccttgt tgactctccc 80 1090 80 DNA Rat 1090ctctcaccgg actcttgtag attcaacttg ccgctgtctt ttaggctttg tacttggctt 60ttccactttc gcactctccc 80

1. A standardizing control for RNA samples to be tested on non-controlgene sequences on nucleic acid arrays, comprising a pool of uniquetagged synthetic antisense mRNA molecules of a known concentration,wherein any two sequences are unique if their sequences differ.
 2. Thestandardizing control of claim 1, wherein said antisense molecules areunique if their sequences differ by about 20%-100% of the nucleic acidsequence from other antisense molecules in the pool.
 3. Thestandardizing control of claim 2 wherein wherein said antisensemolecules are unique if their sequences differ by about 25%-80% of thenucleic acid sequence with the other antisense molecules in the pool. 4.The standardizing control of claim 3, wherein said antisense moleculesare unique if their sequences differ by about 30%-70% of the nucleicacid sequences with the other antisense molecules in the pool.
 5. Thestandardizing control of claim 1, wherein the number of unique taggedantisense molecules in the set of tagged mRNA antisense molecules isabout 50 to 50,000.
 6. The standardizing control of claim 5, wherein thenumber of unique tagged antisense molecules in the set of tagged mRNAantisense molecules is about 100 to 40,000.
 7. The standardizing controlof claim 6, wherein the number of unique tagged antisense molecules inthe pool of tagged mRNA antisense molecules is about 200 to 35,000. 8.The standardizing control of claim 1, wherein the pool includes a taggedantisense nucleic acid for each non-control sequence that may be presentin the sample.
 9. The standardizing control of claim 1, wherein the poolincludes a representative or representational number of tagged antisensemolecules.
 10. The standardizing control of claim 9, wherein theantisense molecules included in the pool includes a representationalnumber of tagged antisense molecules, the total number of differenttagged mRNA antisense molecules in any given set is a fraction of thetotal number of different or distinct mRNAs in the sample employed togenerate an antisense target.
 11. The standardizing control of claim 10,wherein the total number of tagged antisense molecules in the pool willnot exceed about 80% of the total number of distinct mRNAs in theoriginal sample.
 12. The standardizing control of claim 11, wherein thetotal number of tagged antisense molecules in the pool will not exceedabout 60-50% of the total number of distinct mRNAs in the originalsample.
 13. The standardizing control of claim 11, wherein the totalnumber of tagged antisense molecules in the pool will not exceed about40-20% of the total number of distinct mRNAs in the original sample. 14.The standardizing control of claim 1, wherein the non-control genesequences on the nucleic acid array are selected from the groupcomprising oncogenes; genes encoding tumor suppressors; genes encodingcell cycle regulators; stress response genes; genes encoding ion channelproteins; genes encoding transport proteins; genes encodingintracellular signal transduction modulator and effector factors;apoptosis related genes; DNA synthesis/recombination/repair genes; genesencoding transcription factors; genes encoding DNA-binding proteins;genes encoding receptors, and genes encoding cell-cell communicationproteins
 15. The standardizing control of claim 14 wherein the genesencoding receptors are selected from the group comprising receptors forgrowth factors, chemokines, interleukins, interferons, hormones,neurotransmitters, cell surface antigens, and cell adhesion molecules.16. The standardizing control of claim 14 wherein the genes encodingcell-cell communication proteins are selected from the group comprisinggrowth factors, cytokines, chemokines, interleukins, interferons, andhormones.
 17. The standardizing control of claim 1, wherein saidnon-control gene sequences are mammalian.
 18. The standardizing controlof claim 17, wherein said mammalian sequences are selected from thegroup comprising human, rat, mouse, and bovine.
 19. The standardizingcontrol of claim 1, wherein the pool of tagged antisense molecules haveat least two different gene functional classes represented in a givenset.
 20. The standardizing control of claim 19, wherein the number ofdifferent functional classes of genes represented in a given set isabout 2-5.
 21. The standardizing control of claim 20, wherein the numberof different functional classes of genes represented in a given set isabout
 24. 22. The standardizing control of claim 21, wherein the numberof different functional classes of genes represented in a given set isabout 2-3.
 23. A test kit for standardizing results from nucleic acidarrays comprising in a least one container the pool of unique taggedmRNA antisense molecules in claim
 1. 24. The test kit of claim 23,wherein there are different tagged antisense molecules in the pool. 25.The test kit of claim 24, wherein the number of different taggedantisense molecules in the pool is from about 20 to 40,000.
 26. The testkit of claim 25, wherein the number of different tagged antisensemolecules in the pool is from about 20 to 10,000.
 27. The test kit ofclaim 26, wherein the number of different tagged antisense molecules inthe pool is from about 50 to 2,000.
 28. The test kit of claim 27,wherein the number of different tagged antisense molecules in the poolis from about 75 to 1,500.